Cyclic peptide derivative

ABSTRACT

The present invention aims to provide cyclic peptide derivatives having motilin receptor antagonist activity or the like and useful as pharmaceuticals.  
     The present invention provides a compound of general formula ( 1 ):  
                 
 
     wherein R 1  represents an optionally substituted phenyl group or the like; R 2  represents an amino group or the like; R 3  to R 6  represent a hydrogen atom, a methyl group or the like; R 7  represents a hydrogen atom or the like; V to Z represent a carbonyl group or a methylene group; m represents an integer of 0-2; and n represents an integer of 0-3; or a hydrate or a pharmaceutically acceptable salt thereof.

TECHNICAL FIELD

[0001] The present invention relates to cyclic peptide derivativeshaving motilin receptor antagonist activity or the like and useful aspharmaceuticals.

BACKGROUND ART

[0002] Among gastrointestinal hormones, motilin is a straight peptideconsisting of 22 amino acids that is known to control gastrointestinalmotility of mammals including human. Motilin receptors have been knownto be predominantly localized in the upper gastrointestinal tract suchas stomach and duodenum, and recently found to be also localized in thelower gastrointestinal tract such as large intestine (William et al.,Am. J. Physiol., 262, G50-G55 (1992)), showing that motilin may beinvolved in the motility of not only the upper but also the lowergastrointestinal tract.

[0003] It was reported that patients of irritable bowel syndrome showingdiarrhea conditions or patients of irritable bowel syndrome under stressshow hypermotilinemia (Preston et al., Gut, 26, 1059-1064 (1985); Fukodoet al., Tohoku J. Exp. Med., 151, 373-385 (1987)), suggesting thatincreased blood motilin may be involved in this pathology. Otherpathologies reported to be associated with hypermotilinemia includeCrohn's disease, ulcerative colitis, pancreatitis, diabetes, obesity,malabsorption syndrome, bacterial diarrhea, atrophic gastritis,postgastrectomy/enterectomy, etc. Therefore, motilin receptorantagonists may potentially improve pathologies with increased bloodmotilin such as irritable bowel syndrome.

[0004] Recently, efforts have been made to develop and research motilinreceptor antagonists, and various compounds have been reported(JP-A-7-138284, JP-A-2000-44595, etc.).

[0005] Especially, JP-A-7-138284 discloses cyclic peptide derivatives,which are used as pharmacological tools in studies of the effect ofmotilin on gastrointestinal motility or development and research ofpharmaceuticals in this field of the art. However, their motilinantagonist activity is not sufficient and it would be desirable todevelop cyclic peptide derivatives having higher activity.

DISCLOSURE OF THE INVENTION

[0006] An object of the present invention is to provide cyclic peptidederivatives having motilin receptor antagonist activity and useful aspharmaceuticals.

[0007] As a result of careful studies to develop novel cyclic peptidederivatives having higher motilin receptor antagonist activity, weaccomplished the present invention on the basis of the finding thatcyclic peptide derivatives of general formula (1) have excellent motilinreceptor antagonist activity.

[0008] Accordingly, the present invention provides a compound of generalformula (1):

[0009] wherein

[0010] R₁ represents an optionally substituted phenyl group or anoptionally substituted heterocycle;

[0011] R₂ represents a hydrogen atom or an optionally substituted aminogroup;

[0012] R₃ represents a hydrogen atom, a methyl group or an ethyl group;

[0013] R₄ represents a hydrogen atom, a methyl group or an ethyl group;

[0014] R₅ represents a hydrogen atom, a methyl group or an ethyl group;

[0015] R₆ represents a hydrogen atom, a methyl group, an ethyl group or—CO—R₇;

[0016] R₇ represents a hydrogen atom or a straight or branched alkylgroup having 1-3 carbon atoms;

[0017] V represents a carbonyl group or a methylene group;

[0018] W represents a carbonyl group or a methylene group;

[0019] X represents a carbonyl group or a methylene group;

[0020] Y represents a carbonyl group or a methylene group;

[0021] Z represents a carbonyl group or a methylene group;

[0022] m represents a number of 0-2; and

[0023] n represents a number of 0-3;

[0024] except for the case where R₁ represents a phenyl group, and

[0025] R₂ represents an amino group, and all of R₃, R₄, R₅ and R₆represent a hydrogen atom, and V represents a methylene group, and allof W, X, Y and Z represent a carbonyl group, and both m and n represent1; or

[0026] a hydrate or a pharmaceutically acceptable salt thereof.

[0027]

[0028] The present invention also provides a pharmaceutical comprisingthe compound of general formula (1) or a hydrate or a pharmaceuticallyacceptable salt thereof as an active ingredient. The present inventionalso provides a motilin receptor antagonist comprising the abovecompound or a hydrate or a pharmaceutically acceptable salt thereof. Thepresent invention also provides a gastrointestinal motility inhibitorcomprising the above compound or a hydrate or a pharmaceuticallyacceptable salt thereof as an active ingredient. The present inventionalso provides a pharmaceutical for treating hypermotilinemia comprisingthe above compound or a hydrate or a pharmaceutically acceptable saltthereof as an active ingredient. The above compound or a hydrate or apharmaceutically acceptable salt thereof may also be used for preparinga pharmaceutical or a pharmaceutical composition such as motilinreceptor antagonists, gastrointestinal motility depressants andpharmaceuticals for treating hypermotilinemia.

[0029] According to another aspect of the present invention, a methodfor treating hypermotilinemia with the above compound or a hydrate or apharmaceutically acceptable salt thereof is provided. Preferably, amethod for treating hypermotilinemia associated with irritable bowelsyndrome, Crohn's disease, ulcerative colitis, pancreatitis, diabetes,obesity, malabsorption syndrome, bacterial diarrhea, atrophic gastritis,postgastrectomy/enterectomy and the like is provided. For example, amethod comprising the step of administering a therapeutically effectiveamount of the above compound or a hydrate or a pharmaceuticallyacceptable salt thereof to a patient in need of such treatment isprovided.

[0030] The present invention also provides a compound of general formula(2):

[0031] wherein R₃, R₄, R₅, R₆, m and n have the same meanings as definedfor general formula (1) above;

[0032] P₂ represents a hydrogen atom or a protective group for aphenolic hydroxyl group;

[0033] P₃ represents a hydrogen atom or a protective group for acarboxyl group;

[0034] P₄ represents a hydrogen atom or a protective group for an aminogroup; and

[0035] P₅ represents a hydrogen atom or a protective group for an aminogroup; or

[0036] a hydrate or a pharmaceutically acceptable salt thereof.

[0037] The present invention also provides a compound of general formula(3):

[0038] wherein R₃, R₄, R₅, m, n, P₂, P₃ and P₅ have the same meanings asdefined for general formula (2) above;

[0039] R₁ has the same meaning as defined in general formula (1) above;and

[0040] R₂′ represents a hydrogen atom or an optionally substitutedprotected amino group; or

[0041] a hydrate or a pharmaceutically acceptable salt thereof.

[0042] The present invention also provides a compound of general formula(4):

[0043] wherein R₁, R₃, R₄, R₅, R₂′, m and n have the same meanings asdefined for general formula (3) above;

[0044] P₆ represents a hydrogen atom or a protective group for acarboxyl group; and

[0045] P₇ represents a hydrogen atom or a protective group for an aminogroup; or

[0046] a hydrate or a pharmaceutically acceptable salt thereof.

[0047] In the definition of the compound of general formula (1),substituents in the optionally substituted phenyl group for R₁preferably include halogen atoms and trifluoromethyl, hydroxyl, aminoand nitrile groups, more preferably halogen atoms, especially fluorineatom. The phenyl group may have one or more of these substituents whichmay be the same or different. The number of substituents is preferably1-3, more preferably 1.

[0048] The optionally substituted phenyl group for R₁ may be phenyl,4-fluorophenyl, 3-fluorophenyl, 2-fluorophenyl, 3,4-difluorophenyl,2,3,4-trifluorophenyl, 4-chlorophenyl, 3-chlorophenyl, 2-chlorophenyl,3,4-dichlorophenyl, 2,3,4-trichlorophenyl, 4-trifluoromethylphenyl,3-trifluoromethylphenyl, 4-hydroxyphenyl, 3-hydroxyphenyl,4-aminophenyl, 4-cyanophenyl, 3-cyanophenyl, 3-fluoro-4-hydroxyphenyl,2-fluoro-4-hydroxyphenyl or the like, preferably phenyl, 4-fluorophenyl,3-fluorophenyl, 3,4-difluorophenyl, 4-chlorophenyl, 3-chlorophenyl or3,4-dichlorophenyl, more preferably 4-fluorophenyl.

[0049] The heterocycle in the optionally substituted heterocycle for R₁may be an aliphatic or aromatic 5 to 7-membered monocycle or fused cyclecontaining at least one hetero atoms selected from nitrogen, sulfur andoxygen atoms, specifically pyridyl, furyl, thienyl, indolyl, quinolinyl,benzofuryl, tetrahydroisoquinolyl, preferably indolyl.

[0050] Substituents in the optionally substituted heterocycle for R₁include hydroxyl, amino, carboxyl, methoxyl, methyl, ethyl,trifluoromethyl, oxo, etc., and the heterocycle may have one or more ofthese substituents which may be the same or different.

[0051] The optionally substituted heterocycle for R₁ is preferably3-indolyl.

[0052] R₁ as defined above is preferably phenyl, 4-fluorophenyl,3-fluorophenyl, 3,4-difluorophenyl, 4-chlorophenyl, 3-chlorophenyl,3,4-dichlorophenyl or 3-indolyl, more preferably 4-fluorophenyl.

[0053] Substituents in the optionally substituted amino group for R₂ maybe, for example, a straight or branched alkyl group having 1-3 carbonatoms, preferably methyl or ethyl. The amino group may have one or moreof these substituents which may be the same or different.

[0054] The optionally substituted amino group for R₂ may be an aminogroup which may be substituted by one or more the same or differentstraight or branched alkyl groups having 1-3 carbon atoms, such asamino, methylamino, ethylamino, dimethylamino, among which amino isespecially preferred.

[0055] R₂ as defined above is preferably an amino group.

[0056] R₃ is preferably a hydrogen atom or a methyl group.

[0057] R₄ is preferably a hydrogen atom or a methyl group.

[0058] R₅ is preferably a hydrogen atom or a methyl group.

[0059] R₆ is preferably a hydrogen atom, a methyl group or an acetylgroup.

[0060] V is preferably a methylene group.

[0061] W is preferably a carbonyl group or a methylene group.

[0062] X is preferably a carbonyl group or a methylene group.

[0063] Y is preferably a carbonyl group.

[0064] Z is preferably a carbonyl group.

[0065] m is preferably 0 or 1.

[0066] n is preferably 0, 1 or 2.

[0067] The sum of m and n is preferably 1, 2 or 3.

[0068] In a preferred compound of general formula (1), X is a methylenegroup when R₁ a phenyl group and R₃ is a hydrogen atom.

[0069] In another preferred compound of general formula (1), R₁ is a4-fluorophenyl group, R₂ is an amino group, R₃ is a hydrogen atom or amethyl group, R₄ is a hydrogen atom or a methyl group, R₅ is a hydrogenatom or a methyl group, R₆ is a hydrogen atom, a methyl group or anacetyl group, V is a methylene group, W is a carbonyl group or amethylene group, X is a carbonyl group or a methylene group, Y is acarbonyl group, Z is a carbonyl group, m is 0 or 1, n is 0, 1 or 2 andthe sum of m and n is 1, 2 or 3. More preferably, the compound is anyone of(2S-(2S,13S))-2-amino-N-(2-(3-tert-butyl-4-hydroxyphenylmethyl)-1,4,8-triaza-3,7,14-trioxocyclotetradec-13-yl)-3-(4-fluorophenyl)-N-methylpropionamide,(2S-(2S,13S))-2-amino-N-(2-(3-tert-butyl-4-hydroxyphenylmethyl)-1-methyl-1,4,8-triaza-3,7,14-trioxocyclotetradec-13-yl)-3-(4-fluorophenyl)-N-methylpropionamide,(2S-(2S,12S))-2-amino-N-(2-(3-tert-butyl-4-hydroxyphenylmethyl)-1,4,8-triaza-3,7,13-trioxocyclotridec-12-yl)-3-(4-fluorophenyl)-N-methylpropionamide,(2S-(2S,13S))-2-amino-N-(2-(3-tert-butyl-4-hydroxyphenylmethyl)-4-methyl-1,4,8-triaza-3,7,14-trioxocyclotetradec-13-yl)-3-(4-fluorophenyl)-N-methylpropionamide,(2S-(2S,13S))-2-amino-N-(2-(3-tert-butyl-4-hydroxyphenylmethyl)-8-methyl-1,4,8-triaza-3,7,14-trioxocyclotetradec-13-yl)-3-(4-fluorophenyl)-N-methylpropionamide,(2S-(2S,14S))-2-amino-N-(2-(3-tert-butyl-4-hydroxyphenylmethyl)-1,4,9-triaza-3,8,15-trioxocyclopentadec-14-yl)-3-(4-fluorophenyl)-N-methylpropionamide,(2S,14S)-13-(2S-2-amino-3-(4-fluorophenyl)propylamino)-2-(3-tert-butyl-4-hydroxyphenylmethyl)-1,4,8-triaza-3,7,14-trioxocyclotetradecane,(2S-(2S,13S))—N-(8-acetyl-2-(3-tert-butyl-4-hydroxyphenylmethyl)-N-methyl-1,4,8-triaza-3,14-dioxocyclotetradec-13-yl)-2-amino-3-(4-fluorophenyl)propionamide,(2S-(2S,13S))-2-amino-N-(2-(3-tert-butyl-4-hydroxyphenylmethyl)-9,N-dimethyl-1,4,9-triaza-3,8,14-trioxocyclotetradec-13-yl)-3-(4-fluorophenyl)propionamide,and(2S-(2S,13S))-2-amino-N-(2-(3-tert-butyl-4-hydroxyphenylmethyl)-1,4,8-triaza-3,7,14-trioxocyclotetradec-13-yl)-3-phenyl-N-methylpropionamide.

[0070] The compounds of general formulae (2)-(4) are useful asintermediates for preparing the compound of general formula (1).Protective groups for amino, carboxyl and phenolic hydroxyl groups asdefined for these general formulae (2)-(4) include the following groups.

[0071] Protective groups for amino groups (i.e. P₄, P₅, P₇ and theprotective group in R₂% which is an optionally substituted protectedamino group) include functional groups known to be useful as protectivegroups for amino groups such as benzyloxycarbonyl, t-butoxycarbonyl,9-fluorenylmethyloxycarbonyl, allyloxycarbonyl,2,2,2-trichloroethoxycarbonyl, benzoyl, acetyl, trifluoroacetyl,benzenesulfonyl, p-toluenesulfonyl, trimethylsilyl,t-butyldimethylsilyl, benzyl and benzyloxymethyl, preferablybenzyloxycarbonyl and t-butoxycarbonyl.

[0072] Protective groups for carboxyl groups (P₃ and P₆) includefunctional groups known to be useful as protective groups for carboxylgroups such as methyl, ethyl, t-butyl, allyl, benzyl,2,2,2-trichloroethyl, trimethylsilyl and t-butyldimethylsilyl,preferably methyl, benzyl and t-butyl.

[0073] Protective groups for phenolic hydroxyl groups (P₂) includefunctional groups known to be useful as protective groups for phenolichydroxyl groups such as methyl, methoxymethyl, benzyloxymethyl,2-methoxyethoxymethyl, 2-(trimethylsilyl)ethoxymethyl,tetrahydropyranyl, allyl, t-butyl, benzyl, 4-methoxybenzyl,2-nitrobenzyl, trimethylsilyl, t-butyldimethylsilyl, acetyl,trifluoroacetyl, benzoyl, methoxycarbonyl, 2,2,2-trichloroethoxycarbonyland t-butoxycarbonyl, preferably benzyl.

[0074] Acids forming a salt of the compound of formula (1), (2), (3) or(4) include inorganic acids such as hydrochloric acid, hydrobromic acid,hydroiodic acid, sulfuric acid and phosphoric acid, and organic acidssuch as acetic acid, oxalic acid, maleic acid, fumaric acid, citricacid, succinic acid, tartaric acid, methanesulfonic acid andtrifluoroacetic acid.

[0075] Compounds of the present invention may exist as optical isomers,and such individual optical isomers and mixtures thereof are allincluded in the present invention.

[0076] Compounds of the present invention may also be obtained ashydrates of the compounds of formula (1), (2), (3) or (4).

[0077] The present invention is specifically explained below, in whichamino acids forming peptides, protective groups and reagents aresometimes abbreviated as follows. Tyr: tyrosine; Z: benzyloxycarbonyl;Boc: tert-(or t-)butoxycarbonyl; CMPI: 2-chloro-1-methylpyridiniumiodide; PyCIU: chloro-N,N,N′,N′-bis(tetramethylene)formamidiniumhexafluorophosphate; PyBrop: bromotripyrrolidinophosphoniumhexafluorophosphate; BOP:benzotriazole-1-yl-oxy-tris(dimethylamino)phosphoniumhexafluorophosphate; HATU:O-(7-azabenzotriazole-1-yl)-1,1,3,3-tetramethyluroniumhexafluorophosphate; WSCI: N-ethyl-N′-3-dimethylaminopropylcarbodiimide; DIC: N,N′-diisopropyl carbodiimide; DCC: N,N′-dicyclohexylcarbodiimide; DPPA: diphenylphosphoryl azide; CDI:1,1′-carbonyldiimidazole; HOBT: 1-hydroxybenzotriazole monohydrate; NMM:N-methylmorpholine; TEA: triethylamine; DIEA: diisopropylethylamine;DMAP: 4-dimethylaminopyridine; TFA: trifluoroacetic acid; THF:tetrahydrofuran; DMF: N,N-dimethylformamide; CH: chloroform; MC:methylene chloride; M: methanol; N: concentrated aqueous ammonia; EA:ethyl acetate; H: n-hexane.

PREFERRED EMBODIMENTS OF THE INVENTION

[0078] The present application claims priority based on Japanese PatentApplication No. 2000-253950, the disclosure of which is incorporatedherein as reference in its entirety.

[0079] Compounds of the present invention can be prepared by solid phaseor liquid phase processes. In the case of solid phase processes, theycan be prepared by using an automatic organic synthesizer or manually.

[0080] Compounds of the present invention can be prepared by the processof scheme 1, 2 or 3 described below, which may be modified in part asappropriate to suit the desired compound, starting from a knowncompound. Compounds of the present invention can also be obtained byappropriately adapting the specific processes described in the examplesbelow.

[0081] Scheme 1

[0082] Compounds of the present invention wherein V represents amethylene group and W, X, Y and Z represent a carbonyl group can beprepared according to scheme 1 shown below using reagents having desiredgroups.

[0083] wherein P₁, P₄ and P₅ represent a protective group for an aminogroup, P₂ represents a hydrogen atom or a protective group for aphenolic hydroxyl group, P₃ represents a protective group for a carboxylgroup, R₂′ represents a hydrogen atom or an optionally substitutedprotected amino group and the other symbols have the same meanings asdefined in claim 1.

[0084] In scheme 1, conversion from compound 1 (described in Example1(1) of JPA 2000-44595) into compound 2 can be performed by protectingthe amino group of compound 1 and then, if desired, protecting thephenol group. Specifically, compound 2 wherein P₂ is a hydrogen atom canbe obtained by reacting compound 1 with a protective group-introducingreagent such as benzyloxycarbonyl chloride or di-t-butyl dicarbonate ina mixed solvent of dioxane and water in the presence of a base such assodium carbonate with ice cooling or at room temperature. Compound 2wherein P₂ is a protective group (especially, benzyl) can be obtained byfurther reacting the compound with benzyl chloride or the like in DMF orTHF in the presence of a base such as sodium hydride with ice cooling.

[0085] Conversion from compound 2 into compound 3 can be performed bytreating the methyl ester group of compound 2 with a base to hydrolyzeit. Specifically, compound 3 can be obtained by stirring compound 2 witha base such as sodium hydroxide or lithium hydroxide in a mixed solventof methanol and water or dioxane and water with ice cooling or at roomtemperature.

[0086] Conversion from compound 3 into compound 4 can be performed byreacting compound 3 with a methylating agent or an ethylating agent inthe presence of a base. Specifically, compound 4 can be obtained byreacting compound 3 with methyl iodide, dimethyl sulfate or ethyl iodideor the like in DMF or THF or a mixed solvent of DMF and THF in thepresence of a base such as sodium hydride with ice cooling. The phenolichydroxyl group here is pref erably protected by P₂. This methylation orethylation reaction is unnecessary for compounds wherein R₄ is ahydrogen atom.

[0087] Conversion from compound 4 into compound 6 is performed byreacting compound 4 with compound 5. This reaction can be performed bycondensing the carboxyl group of compound 4 with the amino group ofcompound 5 using a known conventional condensing reagent. Specifically,compound 6 can be obtained by stirring a mixture of compound 4, compound5 and one of condensing agents such as CMPI, BOP, PyCIU, PyBrop, HATU,DCC, DIC, WSCI, DPPA and CDI in a reaction solvent such as THF or DMF ormethylene chloride. The reaction here is often preferably promoted byadding a base such as TEA, DIEA, NMM, pyridine and DMAP or adding anadditive such as HOBT as appropriate. Compound 5 may be commerciallyavailable. If not commercially available, compound 5 wherein R₅ is ahydrogen atom can be prepared by esterifying a suitable α-, β-, γ- orδ-amino acid. Compound 5 wherein R₅ is a methyl or ethyl group can beobtained by protecting the amino group of a suitable α-, β-, γ- orδ-amino acid ester with a benzyloxycarbonyl group or a t-butoxycarbonylgroup in the same manner as described for the conversion from compound 1into compound 2, methylating or ethylating the amino acid ester in thesame manner as described for the conversion from compound 3 intocompound 4, and then eliminating the amino-protecting group by aconventional method.

[0088] Conversion from compound 6 into compound 7 can be performed byeliminating the amino-protecting group P₁ of compound 6. A specificdeprotecting method comprises hydrogenolysis using a palladium catalystwhen P₁ is a benzyloxycarbonyl group or treatment with an acid such asTFA when it is a t-butoxycarbonyl group.

[0089] Conversion from compound 7 into compound 9 is performed byreacting compound 7 with compound 8. This reaction can be performed bycondensing the amino group of compound 7 with the carboxyl group ofcompound 8 using a known conventional condensing reagent. Specifically,compound 9 can be obtained by the same procedure as described for theconversion from compound 4 into compound 6. Compound 8 may becommercially available. If not commercially available, it can beobtained by simultaneously methylating both protected amino groups of aprecursor of compound 8 wherein the α-amino group and the pendant aminogroup corresponding to R₃ and R₆ are protected by P₄ and P₅ respectively(P₄ and P₅ are preferably different protective groups) in the samemanner as described for the conversion from compound 3 into compound 4.Compounds 8 other than those wherein both R₃ and R₆ represent a methylgroup can also be prepared by appropriately adapting this procedure.

[0090] Conversion from compound 9 into compound 10 can be performed byeliminating the amino-protecting group P₄ of compound 9. A specificdeprotecting method comprises hydrogenolysis using a palladium catalystwhen P₄ is a benzyloxycarbonyl group or treatment with an acid such asTFA when it is a t-butoxycarbonyl group.

[0091] Conversion from compound 10 into compound 12 is performed byreacting compound 10 with compound 11. This reaction can be performed bycondensing the amino group of compound 10 with the carboxyl group ofcompound 11 using a known conventional condensing reagent. Specifically,compound 12 can be obtained by the same procedure as described for theconversion from compound 4 into compound 6. Compound 11 may becommercially available. If not commercially available, compound 11wherein R₂′ is a protected methylamino group can be obtained bymethylating a precursor of compound 11 wherein the group correspondingto R₂′ is a protected amino group in the same manner as described forthe conversion from compound 3 into compound 4. Compounds 11 other thanthose wherein the group corresponding to R₂′ is a protected amino groupcan also be prepared by appropriately adapting this procedure.

[0092] Conversion from compound 12 into compound 13 can be performed bysimultaneously or stepwise eliminating the amino-protecting group P₅,carboxyl-protecting group P₃ and phenol-protecting group P₂ of compound12. A specific deprotecting method comprises hydrogenolysis using apalladium catalyst when P₅ is a benzyloxycarbonyl group, P₃ is a benzylgroup and P₂ is a benzyl group or treatment with an acid such as TFAwhen P₅ is a t-butoxycarbonyl group and P₃ is t-butyl or alkalihydrolysis when P₃ is a methyl group, for example.

[0093] Conversion from compound 13 into compound 14 can be performed byintramolecular cyclization of the amino group and the carboxyl group ofcompound 13 using a condensing reagent known to be useful for macrocycleformation reaction. Specifically, compound 14 can be obtained bystirring compound 13 at room temperature under dilute conditions (at aconcentration of compound 13 of about 0.005-0.02 M, preferably 0.01 M)using DMF, BOP and pyridine as reaction solvent, condensing agent andbase, respectively.

[0094] When R₂′ of compound 14 is an optionally substituted protectedamino group, conversion from compound 14 into compound 15 can beperformed by deprotecting the amino group. A specific method compriseshydrogenolysis using a palladium catalyst when the amino-protectinggroup of R₂′ is a benzyloxycarbonyl group or treatment with an acid suchas TFA when it is a t-butoxycarbonyl group. When R₂′ is a hydrogen atom,compound 14 is a desired compound.

[0095] Scheme 2

[0096] Compounds of the present invention wherein V and X represent amethylene group and W, Y and Z represent a carbonyl group can beprepared according to scheme 2 shown below using reagents having desiredgroups.

[0097] wherein P₄ and P₅ represent a protective group for an aminogroup, P₂ represents a hydrogen atom or a protective group for aphenolic hydroxyl group, P₃ represents a protective group for a carboxylgroup, R₂′ represents a hydrogen atom or an optionally substitutedprotected amino group when R₂ of the desired compound represents anoptionally substituted amino group, R₇ represents a functional groupcapable of reacting with an amino group to form a bond such as formyl,—CH₂Hal where Hal represents a halogen atom such as a chlorine, bromineor iodine atom, —CH₂OSO₂R where R represents a methyl, trifluoromethyl,tosyl or the like group, and the other symbols have the same meanings asdefined in claim 1.

[0098] Conversion from compound 9 into compound 16 can be performed bysimultaneously or stepwise eliminating the amino-protecting group P₅,carboxyl-protecting group P₃ and phenol-protecting group P₂ of compound9. A specific deprotecting method comprises hydrogenolysis using apalladium catalyst when P₅ is a benzyloxycarbonyl group, P₃ is a benzylgroup and P₂ is a benzyl group or treatment with an acid such as TFAwhen P₅ is a t-butoxycarbonyl group and P₃ is t-butyl or alkalihydrolysis when P₃ is a methyl group, for example.

[0099] Conversion from compound 16 into compound 17 can be performed byintramolecular cyclization of the amino group and the carboxyl group ofcompound 16 using a condensing reagent known to be useful for macrocycleformation reaction. Specifically, compound 17 can be obtained bystirring compound 16 at room temperature under dilute conditions (at aconcentration of compound 16 of about 0.005-0.02 M, preferably 0.01 M)using DMF, BOP and pyridine as reaction solvent, condensing agent andbase, respectively.

[0100] Conversion from compound 17 into compound 18 can be performed byeliminating the amino-protecting group P₄ of compound 17. A specificdeprotecting method comprises hydrogenolysis using a palladium catalystwhen P₄ is a benzyloxycarbonyl group or treatment with an acid such asTFA when it is a t-butoxycarbonyl group.

[0101] Conversion from compound 18 into compound 20 is performed byreacting compound 18 with compound 19. When R₇ of compound 19 is aformyl group, this reaction can be performed by reductively forming abond between this formyl group and the amino group of compound 18,specifically stirring a mixture of compound 18 and compound 19 withsodium cyanoborohydride and acetic acid in a reaction solvent such asmethanol or acetonitrile, for example. When R₇ of compound 19 is —CH₂Halor —CH₂OSO₂R or the like, the reaction can be performed by alkylatingthe amino group of compound 18. Compound 19 here wherein R₇ is a formylgroup and R₂′ is an optionally substituted protected amino group, forexample, can be obtained by condensing the carboxyl group of compound 11used in scheme 1 wherein R₂′ represents an optionally substitutedprotected amino group with N,O-dimethylhydroxylamine to convert it intoan N-methoxy-N-methylcarbamoyl group and then reducing theN-methoxy-N-methylcarbamoyl group with lithium aluminium hydride, forexample.

[0102] When R₂′ of compound 20 is an optionally substituted protectedamino group, conversion from compound 20 into compound 21 can beperformed by deprotecting the amino group, specifically catalytichydrogenation when the amino-protecting group of R₂′ is abenzyloxycarbonyl group or treatment with an acid such as TFA when it isa t-butoxycarbonyl group. When R₂′ is a hydrogen atom, compound 21 is adesired compound.

[0103] Scheme 3

[0104] Compounds of the present invention wherein R₆ represents anacetyl group, V and W represent a methylene group and X, Y and Zrepresent a carbonyl group can be prepared according to scheme 3 shownbelow using reagents having desired groups.

[0105] wherein P₁, P₄, P₅ and P₇ represent a protective group for anamino group, P₂ represents a hydrogen atom or a protective group for aphenolic hydroxyl group, P₆ represents a protective group for a carboxylgroup, R₂′ represents a hydrogen atom or an optionally substitutedprotected amino group when R₂ of the desired compound represents anoptionally substituted amino group, R₈ represents a functional groupcapable of reacting with an amino group to form a bond such as formyl,—CH₂Hal where Hal represents a halogen atom such as a chlorine, bromineor iodine atom, —CH₂OSO₂R where R represents a methyl, trifluoromethyl,tosyl or the like group, and the other symbols have the same meanings asdefined in claim 1.

[0106] Conversion from compound 4 into compound 22 can be performed bysuccessively esterifying the carboxyl group of compound 4 with P₆ andeliminating the amino-protecting group P₁ and phenol-protecting groupP₂. Specifically, the carboxyl group can be esterified with P₆ into abenzyl ester by condensation reaction with benzyl alcohol or into at-butyl ester by a reaction with isobutene and an acid or into a methylester by a reaction with methanol or methylation reaction withdiazomethane. The amino-protecting group can be eliminated by treatmentwith an acid such as TFA when it is a t-butoxycarboxy group orhydrogenolysis using a palladium catalyst when it is a benzyloxycarboxygroup. When the phenol-protecting group P₂ is a t-butyl or benzyl group,P₁ and P₂ can be simultaneously eliminated. Compound 22 wherein R₄ is ahydrogen atom and P₆ is a methyl group is compound 1.

[0107] Conversion from compound 22 (or compound 1) into compound 24 isperformed by reacting compound 22 (or compound 1) with compound 23. Thisreaction can be performed by condensing the amino group of compound 22(or compound 1) with the carboxyl group of compound 23 using a knownconventional condensing reagent. Specifically, compound 24 can beobtained by the same procedure as described for the conversion fromcompound 4 into compound 6 in scheme 1. Compound 23 may be commerciallyavailable. If not commercially available, it can be obtained by adaptingthe process for compound 8 described in scheme 1. Compound 23 wherein mis 1, R₃ is a hydrogen atom or a methyl group, any one of P₄ and P₅ is at-butoxycarbonyl group and the other is a benzyloxycarbonyl group iscommercially available.

[0108] Conversion from compound 24 into compound 25 can be performed byeliminating the amino-protecting group P₄ of compound 24. A specificdeprotecting method comprises hydrogenolysis using a palladium catalystwhen P₄ is a benzyloxycarbonyl group or treatment with an acid such asTFA when it is a t-butoxycarbonyl group.

[0109] Conversion from compound 25 into compound 26 is performed byreacting compound 25 with compound 11 used in scheme 1. This reactioncan be performed by condensing the amino group of compound 25 with thecarboxyl group of compound 11 used in scheme 1 using a knownconventional condensing reagent. Specifically, compound 26 can beobtained by the same procedure as described for the conversion fromcompound 4 into compound 6 in scheme 1.

[0110] Conversion from compound 26 into compound 27 can be performed byeliminating the amino-protecting group P₅ of compound 26. A specificdeprotecting method comprises hydrogenolysis using a palladium catalystwhen P₅ is a benzyloxycarbonyl group or treatment with an acid such asTFA when it is a t-butoxycarbonyl group.

[0111] Conversion from compound 27 into compound 29 is performed byreacting compound 27 with compound 28. When R₈ of compound 28 is aformyl group, this reaction can be performed by reductively forming abond with the amino group of compound 27, specifically stirring amixture of compound 27 and compound 28 with sodium cyanoborohydride andacetic acid in a reaction solvent such as methanol or acetonitrile, forexample. When R₇ of compound 28 is —CH₂Hal or —CH₂OSO₂R or the like, thereaction can be performed by alkylating the amino group of compound 27.Compound 28 here wherein R₈ is a formyl group, for example, can beobtained by reducing the carboxyl group of a precursor of compound 28wherein the group corresponding to R₈ is a carboxyl group with borane orthe like into a hydroxymethyl group and then oxidizing the hydroxymethylgroup into a formyl group by the Swern method or the like, for example.Alternatively, it can also be obtained by condensing the carboxyl groupof a precursor of compound 28 wherein the group corresponding to R₈ is acarboxyl group with N,O-dimethylhydroxylamine to convert it into anN-methoxy-N-methylcarbamoyl group and then reducing theN-methoxy-N-methylcarbamoyl group with lithium aluminium hydride.

[0112] Conversion from compound 29 into compound 30 can be performed byacetylating the secondary amino group of compound 29. Specifically,compound 29 can be reacted with an acetylating agent such as acetylchloride or acetic anhydride in a reaction solvent such as methylenechloride, THF or ethyl acetate in the presence of a base such as TEA,DIEA, pyridine or DMAP, for example.

[0113] Conversion from compound 30 into compound 31 can be performed bysimultaneously or stepwise eliminating the amino-protecting group P₇ andthe carboxyl-protecting group P₆ of compound 30. A specific deprotectingmethod comprises hydrogenolysis using a palladium catalyst when P₇ is abenzyloxycarbonyl group and P₆ is a benzyl group or treatment with anacid such as TFA when P₇ is a t-butoxycarbonyl group and P₆ is t-butylor alkali hydrolysis when P₆ is a methyl group.

[0114] Conversion from compound 31 into compound 32 can be performed byintramolecular cyclization of the amino group and the carboxyl group ofcompound 31 using a condensing reagent known to be useful for macrocycleformation reaction. Specifically, compound 32 can be obtained by, forexample, stirring compound 31 at room temperature under diluteconditions (at a concentration of compound 31 of about 0.005-0.02 M,preferably 0.01 M) using DMF, BOP and pyridine as reaction solvent,condensing agent and base, respectively.

[0115] When R₂′ of compound 32 is an optionally substituted protectedamino group, conversion from compound 32 into compound 33 can beperformed by deprotecting the amino group, specifically hydrogenolysisusing a palladium catalyst when the amino-protecting group of R₂′ is abenzyloxycarbonyl group or treatment with an acid such as TFA when it isa t-butoxycarbonyl group. When R₂′ is a hydrogen atom, compound 32 is adesired compound.

[0116] Alternatively, compounds of the present invention can be preparedby appropriately adapting the specific processes described in theexamples below.

EXAMPLES

[0117] The following examples further illustrate the present inventionwithout, however, limiting the invention thereto. The chemicalstructural formulae of the compounds of the examples are shown in TableA-1 and Table A-2 below. TABLE A-1 Example No. Structural formula 1

2

3

4

5

6

7

8

9

10

[0118] In the following examples, ¹H-NMR and mass spectra were measuredwith the following instruments.

[0119]¹H-NMR: JEOL JNM-EX-270 (270 MHz)

[0120] Mass spectra (FAB-MS): JASCO 70-250SEQ

Example 1 Synthesis of(2S-(2S,13S))-2-amino-N-(2-(3-tert-butyl-4-hydroxyphenylmethyl)-1,4,8-triaza-3,7,14-trioxocyclotetradec-13-yl)-3-(4-fluorophenyl)-N-methylpropionamide

[0121] (1) Synthesis of Boc-Tyr(3-tert-Bu)-OH

[0122] To a mixed solution of 14.7 g (58.5 mmol) of H-Tyr(3-tert-Bu)-OMeand 9.30 g (87.8 mmol) of sodium carbonate in 100 ml of 1,4-dioxane and100 ml of water was added 13.4 g (61.4 mmol) of di(tert-butyl)dicarbonate with ice cooling and the mixture was stirred at roomtemperature for 1 hour. The reaction solution was combined with waterand extracted with ethyl acetate, and then the extract was successivelywashed with saturated aqueous NaHCO₃ solution and saturated aqueoussodium chloride solution. The organic layer was dried over anhydrousmagnesium sulfate, and then the solvent was distilled off under reducedpressure to give 24.2 g of crude Boc-Tyr (3-tert-Bu)-OMe.

[0123] To a mixed solution of 16.5 g of the crude Boc-Tyr(3-tert-Bu)-OMe in 120 ml of methanol and 40 ml of water was added 3.30g (78.8 mmol) of lithium hydroxide monohydrate and the mixture wasstirred at room temperature for 80 minutes. The reaction solution wasneutralized with 2N hydrochloric acid with ice cooling and thenacidified with 10% aqueous citric acid. The reaction solution wasextracted with methylene chloride and dried over anhydrous magnesiumsulfate, and then concentrated under reduced pressure to give 15.0 g(quant.) of the title compound.

[0124] (2) Synthesis of Boc-Tyr (3-tert-Bu)-β-Ala-OBzl

[0125] To a solution of 2.50 g (7.42 mmol) of Boc-Tyr (3-tert-Bu)-OH and2.87 g (8.16 mmol) of H-β-Ala-OBzl p-toluenesulfonate in 22 ml of DMFwere added 1.00 g (7.42 mmol) of HOBt, 0.848 ml (7.42 mmol) of NMM and1.56 g (8.16 mmol) of WSCI with ice cooling, and the mixture was stirredat room temperature for 150 minutes. The reaction solution was combinedwith water and extracted with ethyl acetate, and then the extract wassuccessively washed with saturated aqueous NH₄Cl solution, saturatedaqueous NaHCO₃ solution and saturated aqueous sodium chloride solution.The organic layer was dried over magnesium sulfate, and then the solventwas distilled off under reduced pressure to give 3.57 g (97%) of Boc-Tyr(3-tert-Bu)-β-Ala-OBzl.

[0126]¹H-NMR (CDCl₃): δ 1.38 (9H, s), 1.41 (9H, s), 2.40-2.58 (2H, m),2.84-3.00 (2H, m), 3.38-3.50 (2H, m), 4.14-4.26 (1H, m), 5.00 (1H, brs),5.09 (2H, s), 6.20-6.30 (1H, m), 6.54 (1H, d, J=8.2 Hz), 6.85 (1H, dd,J=1.7, 8.2 Hz), 7.03 (1H, d, J=1.7 Hz), 7.30-7.40 (5H, m).

[0127] (3) Synthesis of H-Tyr (3-tert-Bu)-β-Ala-OBzl

[0128] To a solution of 3.55 g (7.13 mmol) of Boc-Tyr(3-tert-Bu)-β-Ala-OBzl in 30 ml of methylene chloride was added 20 ml ofTFA and the mixture was stirred at room temperature for 40 minutes. Thereaction solution was distilled off under reduced pressure, and theresidue was neutralized with saturated aqueous NaHCO₃ solution,extracted with chloroform and the extract was washed with saturatedaqueous sodium chloride solution. The organic layer was dried overmagnesium sulfate, and then the solvent was distilled off under reducedpressured to give 2.60 g (92%) of H-Tyr (3-tert-Bu)-β-Ala-OBzl.

[0129]¹H-NMR (CDCl₃): δ 1.39 (9H, s), 2.58 (2H, t, J=6.3 Hz), 2.92 (1H,d, J=9.5 Hz), 3.13 (1H, dd, J=4.0, 13.9 Hz), 3.48-3.60 (3H, m), 5.14(2H, s), 6.60 (1H, d, J=8.2 Hz), 6.88 (1H, dd, J=2.0, 8.2 Hz), 7.07 (1H,d, J=2.0 Hz), 7.30-7.40 (5H, m), 7.58-7.66 (1H, m).

[0130] (4) Synthesis of Boc-N-Me-Lys (Z)-Tyr (3-tert-Bu)-β-Ala-OBzl

[0131] To a solution of 2.10 g (5.28 mmol) of H-Tyr(3-tert-Bu)-β-Ala-OBzl and 3.19 g (5.54 mmol) of Boc-N-Me-Lys (Z)-OHdicyclohexylammonium salt in 22 ml of DMF were added 713 mg (5.28 mmol)of HOBt, 0.60 ml (5.25 mmol) of NMM and 1.06 g (5.54 mmol) of WSCI withice cooling, and the mixture was stirred at room temperature for 2hours. The reaction solution was combined with water and extracted withethyl acetate, and then the extract was successively washed with 10%aqueous citric acid solution, saturated aqueous NaHCO₃ solution andsaturated aqueous sodium chloride solution. The organic layer was driedover magnesium sulfate, and then the solvent was distilled off underreduced pressure to give 3.42 g (84%) of Boc-N-Me-Lys (Z)-Tyr(3-tert-Bu)-β-Ala-OBzl.

[0132]¹H-NMR (CDCl₃): δ 1.14-1.60 (5H, m), 1.37 (9H, s), 1.45 (9H, s),1.72-1.90 (1H, m), 2.40-2.54 (2H, m), 2.55 (3H, s), 2.86-3.02 (2H, m),3.06-3.22 (2H, m), 3.36-3.50 (2H, m), 4.34-4.56 (2H, m), 4.76-4.88 (1H,m), 5.09 (4H, s), 5.31 (1H, s), 6.30-6.60 (2H, m), 6.58 (1H, d, J=8.2Hz), 6.83 (1H, dd, J=1.9, 8.2 Hz), 7.04 (1H, d, J=1.9 Hz), 7.30-7.42(10H, m).

[0133] (5) Synthesis of N-Me-Lys (Z)-Tyr (3-tert-Bu)-β-Ala-OBzl

[0134] To a solution of 3.10 g (4.01 mmol) of Boc-N-Me-Lys (Z)-Tyr(3-tert-Bu)-β-Ala-OBzl in 20 ml of methylene chloride was added 10 ml ofTFA and the mixture was stirred at room temperature for 90 minutes. Thereaction solution was distilled off under reduced pressure, and theresidue was neutralized with saturated aqueous NaHCO₃ solution,extracted with chloroform and the extract was washed with saturatedaqueous sodium chloride solution. The organic layer was dried overmagnesium sulfate, and then the solvent was distilled off under reducedpressure to give 2.59 g (96%) of N-Me-Lys (Z)-Tyr(3-tert-Bu)-β-Ala-OBzl.

[0135]¹H-NMR (CDCl₃): δ 0.80-1.40 (6H, m), 1.38 (9H, s), 2.30 (3H, s),2.56 (2H, t, J=6.3 Hz), 2.70-3.24 (5H, m), 3.53 (2H, q, J=6.3 Hz),4.48-4.60 (1H, m), 4.80-4.90 (1H, m), 5.12 (2H, s), 5.15 (2H, s),6.62-6.86 (3H, m), 7.04 (1H, d, J=1.3 Hz), 7.30-7.42 (10H, m).

[0136] (6) Synthesis of Boc-Phe (4-F)-N-Me-Lys (Z)-Tyr(3-tert-Bu)-β-Ala-OBzl

[0137] To a solution of 2.50 g (3.71 mmol) of N-Me-Lys (Z)-Tyr(3-tert-Bu)-β-Ala-OBzl and 1.26 g (4.48 mmol) of Boc-Phe (4-F)-OH in18.5 ml of THF were added 1.14 g (4.45 mmol) of CMPI and 1.29 ml (9.28mmol) of TEA with ice cooling and the mixture was stirred at roomtemperature for 3 hours. The reaction solution was combined with waterand extracted with ethyl acetate, and then the extract was washed withsaturated aqueous sodium chloride solution. The organic layer was driedover magnesium sulfate, and then the solvent was distilled off underreduced pressure, and the residue was purified by silica gel columnchromatography (developing solvent; ethyl acetate:hexane=1:1) to give3.48 g (100%) of Boc-Phe (4-F)-N-Me-Lys (Z)-Tyr (3-tert-Bu)-β-Ala-OBzl.

[0138]¹H-NMR (CDCl₃): δ 0.80-1.40 (6H, m), 1.33, 1.36 and 1.39 (total18H, s), 2.17 and 2.61 (total 3H, s), 2.44-3.60 (10H, m), 4.30-4.94 (3H,m), 5.01, 5.04 and 5.10 (total 4H, s), 5.20-5.48 (1H, m), 6.28-7.20(10H, m), 7.30-7.42 (10H, m).

[0139] (7) Synthesis of(2S-(2S,13S))-2-tert-butoxycarbonylamino-N-(2-(3-tert-butyl-4-hydroxyphenylmethyl)-1,4,8-triaza-3,7,14-trioxocyclotetradec-13-yl)-3-(4-fluorophenyl)-N-methylpropionamide

[0140] To a solution of 2.72 g (2.90 mmol) of Boc-Phe (4-F)-N-Me-Lys(Z)-Tyr (3-tert-Bu)-β-Ala-OBzl in 50 ml of methanol was added 270 mg of10% palladium on carbon, and the mixture was stirred overnight at roomtemperature under a hydrogen atmosphere. After filtration, the filtratewas concentrated under reduced pressure to give 2.02 g (97%) of Boc-Phe(4-F)-N-Me-Lys-Tyr (3-tert-Bu)-β-Ala-OH.

[0141] To a mixed solution of 2.02 g (2.83 mmol) of the Boc-Phe(4-F)-N-Me-Lys-Tyr (3-tert-Bu)-β-Ala-OH in 140 ml of DMF and 140 ml ofpyridine was added 3.13 g (7.08 mmol) of BOP reagent and the mixture wasstirred at room temperature for 18 hours. The reaction solution wascombined with water and extracted with ethyl acetate, and the extractwas successively washed with saturated aqueous NH₄Cl solution, saturatedaqueous NaHCO₃ solution and saturated aqueous sodium chloride solution.The organic layer was dried over magnesium sulfate, and then the solventwas distilled off under reduced pressure, and the residue was purifiedby silica gel column chromatography (developing solvent;chloroform:methanol=20:1) to give 1.37 g (69%) of(2S-(2S,13S))-2-tert-butoxycarbonylamino-N-(2-(3-tert-butyl-4-hydroxyphenylmethyl)-1,4,8-triaza-3,7,14-trioxocyclotetradec-13-yl)-3-(4-fluorophenyl)-N-methylpropionamide.

[0142]¹H-NMR (CDCl₃): δ 1.00-2.02 (6H, m), 1.33, 1.37 and 1.39 (total18H, s), 2.30 (3H, s), 2.24-3.86 (10H, m), 4.40-4.90 (4H, m), 5.38-5.60(2H, m), 6.00-6.20 (1H, m), 6.60-7.18 (7H, m), 6.54-7.22 (7H, m).

[0143] (8) Synthesis of(2S-(2S,13S))-2-amino-N-(2-(3-tert-butyl-4-hydroxyphenylmethyl)-1,4,8-triaza-3,7,14-trioxocyclotetradec-13-yl)-3-(4-fluorophenyl)-N-methylpropionamide

[0144] To a solution of 1.25 g (1.79 mmol) of(2S-(2S,13S))-2-tert-butoxycarbonylamino-N-(2-(3-tert-butyl-4-hydroxyphenylmethyl)-1,4,8-triaza-3,7,14-trioxocyclotetradec-13-yl)-3-(4-fluorophenyl)-N-methylpropionamidein 20 ml of methylene chloride was added 10 ml of TFA and the mixturewas stirred at room temperature for 40 minutes. The reaction solutionwas distilled off under reduced pressure, and the residue wasneutralized with saturated aqueous NaHCO₃ solution, extracted withchloroform and the extract was washed with saturated aqueous sodiumchloride solution. The organic layer was dried over magnesium sulfate,and then the solvent was distilled off under reduced pressure, and theresidue was purified by silica gel column chromatography (developingsolvent; chloroform:methanol=20:1) to give 657 mg (61%) of the titlecompound.

[0145] FAB-MS (M⁺+1): 598.

[0146]¹H-NMR (DMSO-d): δ 0.60-1.92 (6H, m), 1.27 and 1.28 (total 9H, s),2.06-2.26 (2H, m), 2.54-2.90 (4H, m), 2.65 (3H, s), 3.06-3.60 (4H, m),3.62-4.94 (3H, m), 6.54-7.22 (7H, m), 7.50-8.50 (3H, m), 9.01 and 9.10(total 1H, s).

Example 2 Synthesis of(2S-(2S.13S))-2-amino-N-(2-(3-tert-butyl-4-hydroxyphenylmethyl)-1-methyl-1.4.8-triaza-3.7.14-trioxocyclotetradec-13-yl)-3-(4-fluorophenyl)-N-methylpropionamide

[0147] (1) Synthesis of Boc-Phe (4-benzyloxy-3-tert-Bu)-OMe

[0148] A suspension of 7.5 g of the crude Boc-Tyr (3-tert-Bu)-OMeobtained in Example 1 (1) and 8.85 g.(64.0 mmol) of potassium carbonatein 45 ml of DMF was stirred at room temperature for 2.5 hours, and then4.6 ml (38.3 mmol) of benzyl bromide was added. The mixture was stirredovernight and then combined with water and extracted with ethyl acetate.The organic layer was washed with saturated aqueous sodium chloridesolution and dried over anhydrous magnesium sulfate and thenconcentrated under reduced pressure, and the residue was purified bysilica gel column chromatography (developing solvent; ethylacetate:n-hexane=1:5) to give 7.5 g (94%) of the title compound.

[0149] (2) Synthesis of Boc-N-Me-Phe (4-benzyloxy-3-tert-Bu)-OH

[0150] To a mixed solution of 7.5 g (17.0 mmol) of Boc-Phe(4-benzyloxy-3-tert-Bu)-OMe in 60 ml of methanol and 20 ml of water wasadded 1.07 g (25.5 mmol) of lithium hydroxide monohydrate, and themixture was stirred at room temperature for 1.5 hours. The reactionsolution was neutralized with 2N hydrochloric acid with ice cooling, andthen acidified with 10% aqueous citric acid. The reaction solution wasextracted with methylene chloride and dried over anhydrous magnesiumsulfate, and then concentrated under reduced pressure to give 6.8 g(93%) of crude Boc-Phe (4-benzyloxy-3-tert-Bu)-OH.

[0151] To a mixed solution of 6.2 g (14.5 mmol) of the crude compound in30 ml of THF and 3 ml of DMF were added 7.2 ml (116 mmol) of methyliodide with ice cooling, then 1.74 g (43.5 mmol) of 60% sodium hydride.The mixture was stirred at room temperature for 21 hours, and thenstirred with 0.58 g (14.5 mmol) of 60% sodium hydride and 2.7 ml (43.5mmol) of methyl iodide for 7 hours. The reaction solution was combinedwith water with ice cooling and washed with a mixed solution of n-hexane(150 ml)-ether (50 ml). The aqueous layer was neutralized with 2Nhydrochloric acid with ice cooling, acidified with 10% aqueous citricacid solution, and then extracted with ethyl acetate. The extract wassuccessively washed with water and saturated aqueous sodium chloridesolution, and then dried over anhydrous magnesium sulfate andconcentrated under reduced pressure to give 5.0 g (78%) of a crudeproduct of the title compound.

[0152] (3) Synthesis of Boc-N-Me-Phe (4-benzyloxy-3-tert-Bu)-β-Ala-OBzl

[0153] To a solution of 2.68 g (6.08 mmol) of the crude Boc-N-Me-Phe(4-benzyloxy-3-tert-Bu)-OH and 2.35 g (6.69 mmol) of H-β-Ala-OBzlp-toluenesulfonate in 20 ml of DMF were added 0.82 g (6.08 mmol) ofHOBt, 0.69 ml (6.08 mmol) of NMM and 1.28 g (6.69 mmol) of WSCI with icecooling, and the mixture was stirred overnight at room temperature. Thereaction solution was combined with water and then 10% aqueous citricacid solution, and then extracted with ethyl acetate. The organic layerwas washed with saturated aqueous NaHCO₃ solution and saturated aqueoussodium chloride solution and dried over magnesium sulfate, and then thesolvent was distilled off under reduced pressure to give 3.70 g (quant.)of the title compound.

[0154] (4) Synthesis of N-Me-Phe (4-benzyloxy-3-tert-Bu)-β-Ala-OBzl

[0155] To a solution of 3.64 g (6.03 mmol) of Boc-N-Me-Phe(4-benzyloxy-3-tert-Bu)-β-Ala-OBzl in 20 ml of methylene chloride wasadded 10 ml of TFA, and the mixture was stirred at room temperature for45 minutes. The reaction solution was concentrated under reducedpressure, and the residue was neutralized with saturated aqueous NaHCO₃solution, and then extracted with methylene chloride and the extract waswashed with saturated aqueous sodium chloride solution. The organiclayer was dried over magnesium sulfate, and then the solvent wasdistilled off under reduced pressure to give 3.02 g (99%) of the titlecompound.

[0156] (5) Synthesis of Boc-N-Me-Lys (Z)-N-Me-Phe(4-benzyloxy-3-tert-Bu)-β-Ala-OBzl

[0157] To a solution of 3.00 g (5.96 mmol) of N-Me-Phe(4-benzyloxy-3-tert-Bu)-β-Ala-OBzl, 4.12 g (7.16 mmol) of Boc-N-Me-Lys(Z)-OH dicyclohexylamine salt and 1.98 g (7.75 mmol) of CMPI in 30 ml ofTHF was added 2.91 ml (20.9 mmol) of TEA, and the mixture was stirredovernight at room temperature. The reaction solution was combined withwater and extracted with ethyl acetate, and the extract was successivelywashed with 10% aqueous citric acid solution, saturated aqueous NaHCO₃solution and saturated aqueous sodium chloride solution. The organiclayer was dried over magnesium sulfate, and then concentrated underreduced pressure, and the residue was purified by silica gel columnchromatography (developing solvent; ethyl acetate:n-hexane=1:1) to give2.95 g (56%) of the title compound.

[0158] (6) Synthesis of Boc-Phe (4-F)-N-Me-Lys (Z)-N-Me-Phe(4-benzyloxy-3-tert-Bu)-β-Ala-OBzl

[0159] To a solution of 2.90 g (3.30 mmol) of Boc-N-Me-Lys (Z)-N-Me-Phe(4-benzyloxy-3-tert-Bu)-β-Ala-OBzl in 20 ml of methylene chloride wasadded 10 ml of TFA, and the mixture was stirred at room temperature for45 minutes. The reaction solution was concentrated under reducedpressure, and the residue was neutralized with saturated aqueous NaHCO₃solution, and then extracted with methylene chloride. The organic layerwas dried over magnesium sulfate, and then the solvent was distilled offunder reduced pressure to give 2.53 g (98%) of crude N-Me-Lys(Z)-N-Me-Phe (4-benzyloxy-tert-Bu)-β-Ala-OBzl.

[0160] To a solution of 2.50 g (3.21 mmol) of the crude compound, 1.00 g(3.53 mmol) of Boc-Phe (4-F)-OH and 0.98 g (3.85 mmol) of CMPI in 16 mlof THF was added 0.98 ml (7.06 mmol) of TEA with ice cooling, and themixture was stirred overnight at room temperature. The reaction solutionwas combined with water and extracted with ethyl acetate, and theextract was successively washed with 10% aqueous citric acid solution,saturated aqueous NaHCO₃ solution and saturated aqueous sodium chloridesolution. The organic layer was dried over magnesium sulfate, and thenconcentrated under reduced pressure, and the residue was purified bysilica gel column chromatography (developing solvent; ethylacetate:hexane=2:1) to give 3.00 g (89%) of the title compound.

[0161] (7) Synthesis of(2S-(2S,13S))-2-tert-butoxycarbonylamino-N-(2-(3-tert-butyl-4-hydroxyphenylmethyl)-1-methyl-1,4,8-triaza-3,7,14-trioxocyclotetradec-13-yl)-3-(4-fluorophenyl)-N-methylpropionamide

[0162] To a solution of 2.72 g (2.60 mmol) of Boc-Phe (4-F)-pN-Me-Lys(Z)-N-Me-Phe (4-benzyloxy-3-tert-Bu)-β-Ala-OBzl in 50 ml of methanol wasadded 0.60 g of palladium hydroxide on carbon, and the mixture wasstirred overnight at room temperature under a hydrogen atmosphere. Afterthe reaction solution was filtered, the filtrate was concentrated underreduced pressure to give 1.98 g of crude Boc-Phe (4-F)-N-Me-Lys-N-Me-Tyr(3-tert-Bu)-β-Ala-OH.

[0163] To a mixed solution of 1.98 g (2.60 mmol) of the crude compoundin 130 ml of DMF and 130 ml of pyridine was added 3.45 g (7.80 mmol) ofBOP reagent, and the mixture was stirred at room temperature for 26hours. The reaction solution was concentrated under reduced pressure,combined with water and extracted with ethyl acetate. The organic layerwas successively washed with 10% aqueous citric acid solution, saturatedaqueous NaHCO₃ solution and saturated aqueous sodium chloride solution,and then dried over magnesium sulfate and concentrated under reducedpressure. The residue was purified by silica gel column chromatography(developing solvent; methylene chloride:methanol:aqueousammonia=20:1:0.1) to give 0.66 g (36%, 2 steps) of the title compound.

[0164] (8) Synthesis of(2S-(2S,13S))-2-amino-N-(2-(3-tert-butyl-4-hydroxyphenylmethyl)-1-methyl-1,4,8-triaza-3,7,14-trioxocyclotetradec-13-yl)-3-(4-fluorophenyl)-N-methylpropionamide

[0165] To a solution of 0.60 g (0.843 mmol) of(2S-(2S,13S))-2-tert-butoxycarbonylamino-N-(2-(3-tert-butyl-4-hydroxyphenylmethyl)-1-methyl-1,4,8-triaza-3,7,14-trioxocyclotetradec-13-yl)-3-(4-fluorophenyl)-N-methylpropionamidein 10 ml of methylene chloride was added 3 ml of TFA, and the mixturewas stirred at room temperature for 1 hour. The reaction solution wasconcentrated under reduced pressure, and the residue was neutralizedwith saturated aqueous NaHCO₃ solution and extracted with methylenechloride. The organic layer was dried over magnesium sulfate, and thenthe solvent was distilled off under reduced pressure, and the residuewas dissolved in 8 ml of methanol with heating. Crystals precipitated atroom temperature were filtered off to give 265 mg (51%) of the titlecompound.

[0166] FAB-MS (M⁺+1): 612.

[0167]¹H-NMR (DMSO-d): δ 1.0-1.5 (6H, m), 1.28 (9H, s), 2.0-2.3 (3H, m),2.4-2.7 (2H, m), 2.48 and 2.64 (total 3H, s), 2.9-3.1 (4H, m), 3.5-3.7(2H, m), 5.22 (1H, d, J=10.9 Hz), 5.33 (1H, t, J=7.3 Hz), 6.5-6.7 (2H,m), 6.9-7.1 (4H, m), 7.1-7.2 (2H, m), 7.7-7.8 (1H, m), 8.97 (1H, s).

[0168] Example 3

(2S-(2S,12S))-2-amino-N-(2-(3-tert-butyl-4-hydroxyphenylmethyl)-1,4,8-triaza-3,7,13-trioxocyclotridec-12-yl)-3-(4-fluorophenyl)-N-methylpropionamide

[0169] (1) Synthesis of Boc-Tyr (3-tert-Bu)-Gly-OBzl

[0170] To a solution of 2.30 g (6.81 mmol) of Boc-Tyr (3-tert-Bu)-OH and1.51 g (7.50 mmol) of H-Gly-OBzl hydrochloride in 20 ml of DMF wereadded 0.92 g (6.81 mmol) of HOBt, 0.78 ml (6.81 mmol) of NMM and 1.44 g(7.50 mmol) of WSCI with ice cooling, and the mixture was stirred atroom temperature for 105 minutes. The reaction solution was combinedwith water and extracted with ethyl acetate, and then the extract wassuccessively washed with 10% aqueous citric acid solution, saturatedaqueous NaHCO₃ solution and saturated aqueous sodium chloride solution.The organic layer was dried over magnesium sulfate, and then the solventwas distilled off under reduced pressure, and the residue was purifiedby silica gel column chromatography (developing solvent; ethylacetate:n-hexane=1:2) to give 1.57 g (48%) of the title compound.

[0171] (2) Synthesis of Boc-N-Me-Lys (Z)-Tyr (3-tert-Bu)-Gly-OBzl

[0172] To a solution of 1.52 g (3.14 mmol) of Boc-Tyr(3-tert-Bu)-Gly-OBzl in 25 ml of methylene chloride was added 8 ml ofTFA, and the mixture was stirred at room temperature for 75 minutes. Thereaction solution was concentrated under reduced pressure, and theresidue was neutralized with saturated aqueous NaHCO₃ solution, and thenextracted with methylene chloride, and the extract was washed withsaturated aqueous sodium chloride solution. The organic layer was driedover magnesium sulfate, and then the solvent was distilled off underreduced pressure to give 1.19 g (98%) of crude H-Tyr(3-tert-Bu)-Gly-OBzl.

[0173] To a solution of 1.15 g (2.99 mmol) of the crude compound and1.89 g (3.29 mmol) of Boc-N-Me-Lys (Z)-OH dicyclohexylamine salt in 20ml of DMF were added 0.40 g (2.99 mmol) of HOBt, 0.34 ml (2.99 mmol) ofNMM and 0.63 g (3.29 mmol) of WSCI with ice cooling, and the mixture wasstirred at room temperature for 2.5 hours. The reaction solution wascombined with water and extracted with ethyl acetate, and then theextract was successively washed with 10% aqueous citric acid solution,saturated aqueous NaHCO₃ solution and saturated aqueous sodium chloridesolution. The organic layer was dried over magnesium sulfate, and thenthe solvent was distilled off under reduced pressure, and the residuewas purified by silica gel column chromatography (developing solvent;ethyl acetate:n-hexane=2:1) to give 2.16 g (95%) of the title compound.

[0174] (3) Synthesis of Boc-Phe (4-F)-N-Me-Lys (Z)-Tyr(3-tert-Bu)-Gly-OBzl

[0175] To a solution of 2.12 g (2.79 mmol) of Boc-N-Me-Lys (Z)-Tyr(3-tert-Bu)-Gly-OBzl in 24 ml of methylene chloride was added 8 ml ofTFA, and the mixture was stirred at room temperature for 1 hour. Thereaction solution was concentrated under reduced pressure, and theresidue was neutralized with saturated aqueous NaHCO₃ solution, and thenextracted with methylene chloride. The organic layer was dried overmagnesium sulfate, and then the solvent was distilled off under reducedpressure to give 1.81 g of crude N-Me-Lys (Z)-Tyr (3-tert-Bu)-Gly-OBzl.

[0176] To a solution of 1.79 g (2.71 mmol) of the crude compound, 0.84 g(2.98 mmol) of Boc-Phe (4-F)-OH and 0.83 g (3.25 mmol) of CMPI in 13 mlof THF was added 0.83 ml (5.96 mmol) of TEA with ice cooling, and themixture was stirred overnight at room temperature. The reaction solutionwas combined with water and extracted with ethyl acetate, and then theextract was successively washed with 10% aqueous citric acid solution,saturated aqueous NaHCO₃ solution and saturated aqueous sodium chloridesolution. The organic layer was dried over magnesium sulfate, and thenconcentrated under reduced pressure, and the residue was purified bysilica gel column chromatography (developing solvent; methylenechloride:methanol:aqueous ammonia=50:1:0.05) to give 2.34 g (90%, 2steps) of the title compound.

[0177] (4) Synthesis of(2S-(2S,12S))-2-tert-butoxycarbonylamino-N-(2-(3-tert-butyl-4-hydroxyphenylmethyl)-1,4,8-triaza-3,7,13-trioxocyclotridec-12-yl)-3-(4-fluorophenyl)-N-methylpropionamide

[0178] To a solution of 1.99 g (2.15 mmol) of Boc-Phe (4-F)-pN-Me-Lys(Z)-Tyr (3-tert-Bu)-Gly-OBzl in 40 ml of methanol was added 0.40 g ofpalladium hydroxide on carbon, and the mixture was stirred overnight atroom temperature under a hydrogen atmosphere. The reaction solution wasfiltered, and then the filtrate was concentrated under reduced pressureto give 1.60 g of crude Boc-Phe (4-F)-N-Me-Lys-Tyr (3-tert-Bu)-Gly-OH.

[0179] To a mixed solution of 1.60 g (2.15 mmol) of the crude compoundin 100 ml of DMF and 100 ml of pyridine was added 2.85 g (6.45 mmol) ofBOP reagent, and the mixture was stirred at room temperature for 23hours. The reaction solution was concentrated under reduced pressure,combined with water, and extracted with ethyl acetate. The organic layerwas successively washed with 10% aqueous citric acid solution andsaturated aqueous sodium chloride solution, and then dried overmagnesium sulfate, and concentrated under reduced pressure. The residuewas purified by silica gel column chromatography (developing solvent;methylene chloride:methanol:aqueous ammonia=25:1:0.05) to give 0.88 g(60%, 2 steps) of the title compound.

[0180] (5) Synthesis of(2S-(2S,12S))-2-amino-N-(2-(3-tert-butyl-4-hydroxyphenylmethyl)-1,4,8-triaza-3,7,13-trioxocyclotridec-12-yl)-3-(4-fluorophenyl)-N-methylpropionamide

[0181] To a solution of 0.86 g (1.26 mmol) of(2S-(2S,12S))-2-tert-butoxycarbonylamino-N-(2-(3-tert-butyl-4-hydroxyphenylmethyl)-1,4,8-triaza-3,7,13-trioxocyclotridec-12-yl)-3-(4-fluorophenyl)-N-methylpropionamidein 15 ml of methylene chloride was added 5 ml of TFA, and the mixturewas stirred at room temperature for 1 hour. The reaction solution wasconcentrated under reduced pressure, and the residue was neutralizedwith aqueous ammonia and extracted with methylene chloride. The organiclayer was dried over magnesium sulfate, and then the solvent wasdistilled off under reduced pressure, and the residue was heated with 4ml of methanol. After standing at room temperature, solids were filteredoff to give 474 mg (65%) of the title compound.

[0182] FAB-MS (M⁺+1): 584.

[0183]¹H-NMR (DMSO-d): δ 0.6-2.0 (6H, m), 1.30 (9H, m), 2.4-3.0 (6H, m),2.58 and 2.78 (total 3H, s), 3.2-3.5 (2H, m), 3.6-4.4 (3.5H, m), 4.9-5.0(0.5H, m), 6.6-7.2 (8H, m), 8.6-8.8 (1H, m), 8.87 (0.5H, d, J=7.3 Hz),9.00 and 9.05 (total 1H, s), 9.17 (0.5H, d, J=7.3 Hz).

Example 4 Synthesis of(2S-(2S,13S))-2-amino-N-(2-(3-tert-butyl-4-hydroxyphenylmethyl)-4-methyl-1.4.8-triaza-3,7,14-trioxocyclotetradec-13-yl)-3-(4-fluorophenyl)-N-methylpropionamide

[0184] (1) Synthesis of Boc-Tyr (3-tert-Bu)-N-Me-β-Ala-OBzl To asolution of 2.41 g (7.15 mmol) of Boc-Tyr (3-tert-Bu)-OH and 1.66 g(8.58 mmol) of N-Me-β-Ala-OBzl (Chem. Pharm. Bull., 31, 10, 1983,3553-3561) in 22 ml of DMF were added 966 mg (7.15 mmol) of HOBt, 0.817ml (7.15 mmol) of NMM and 1.51 g (7.87 mmol) of WSCI with ice cooling,and the mixture was stirred at room temperature for 150 minutes. Thereaction solution was combined with water and extracted with ethylacetate, and then the extract was washed with saturated aqueous NaHCO₃solution and saturated aqueous sodium chloride solution. The organiclayer was dried over magnesium sulfate, and then the solvent wasdistilled off under reduced pressure to give 3.52 g (96%) of Boc-Tyr(3-tert-Bu)-N-Me-β-Ala-OBzl.

[0185]¹H-NMR (CDCl₃): δ 1.34, 1.36, 1.37, 1.40 and 1.42 (total 18H, s),2.00-3.60 (9H, m), 4.66-4.80 (1H, m), 5.09 and 5.10 (total 2H, s),5.16-5.44 (2H, m), 6.40-7.04 (3H, m), 7.30-7.42 (5H, m).

[0186] (2) Synthesis of H-Tyr (3-tert-Bu)-N-Me-β-Ala-OBzl

[0187] To a solution of 3.44 g (6.72 mmol) of Boc-Tyr(3-tert-Bu)-N-Me-β-Ala-OBzl in 30 ml of methylene chloride was added 10ml of TFA, and the mixture was stirred at room temperature for 50minutes. The reaction solution was distilled off under reduced pressure,and the residue was neutralized with saturated aqueous NaHCO₃ solution,extracted with chloroform and the extract was washed with saturatedaqueous sodium chloride solution. The organic layer was dried overmagnesium sulfate, and then the solvent was distilled off under reducedpressure to give 2.70 g (97%) of H-Tyr (3-tert-Bu)-N-Me-β-Ala-OBzl.

[0188]¹H-NMR (CDCl₃): δ 1.37 (9H, s), 2.16-3.14 (4H, m), 2.73 and 2.84(total 3H, s), 3.40-3.70 (2H, m), 3.85 and 3.97 (total 1H, t, J=6.9 Hz),5.08 and 5.10 (total 2H, s), 6.53 and 6.57 (total 1H, d, J=8.3 Hz), 6.80(1H, dd, J=1.3, 8.3 Hz), 6.99 (1H, d, J=1.3 Hz), 7.30-7.44 (5H, m).

[0189] (3) Synthesis of Boc-N-Me-Lys (Z)-Tyr (3-tert-Bu)-N-Me-β-Ala-OBzl

[0190] To a solution of 2.50 g (6.07 mmol) of H-Tyr(3-tert-Bu)-N-Me-β-Ala-OBzl and 3.67 g (6.37 mmol) of Boc-N-Me-Lys(Z)-OH dicyclohexylamine salt in 18 ml of DMF were added 820 mg (6.07mmol) of HOBt, 0.728 ml (6.37 mmol) of NMM and 1.28 g (6.68 mmol) ofWSCI with ice cooling, and the mixture was stirred at room temperaturefor 2 hours. The reaction solution was combined with water and extractedwith ethyl acetate, and then the extract was successively washed withsaturated aqueous NaHCO₃ solution and saturated aqueous sodium chloridesolution. The organic layer was dried over magnesium sulfate, and thenthe solvent was distilled off under reduced pressure to give 4.17 g(87%) of Boc-N-Me-Lys (Z)-Tyr (3-tert-Bu)-N-Me-β-Ala-OBzl.

[0191]¹H-NMR (CDCl₃): δ 1.20-1.60 (5H, m), 1.36 (9H, s), 1.46 (9H, s),1.78-1.92 (1H, m), 2.10-2.94 (4H, m), 2.61, 2.71 and 2.82 (total 6H, s),3.17 (2H, q, J=6.6 Hz), 4.28-4.64 (2H, m), 4.40-5.48 (3H, m), 5.09 and5.10 (total 4H, s), 6.50 and 6.57 (total 1H, d, J=7.9-8.2 Hz), 6.68 (1H,brs), 6.80-6.90 (1H, m), 7.01 (1H, s), 7.28-7.42 (10H, m).

[0192] (4) Synthesis of N-Me-Lys (Z)-Tyr (3-tert-Bu)-N-Me-β-Ala-OBzl

[0193] To a solution of 4.02 g (5.10 mmol) of Boc-N-Me-Lys (Z)-Tyr(3-tert-Bu)-N-Me-β-Ala-OBzl in 25 ml of methylene chloride was added 10ml of TFA, and the mixture was stirred at room temperature for 40minutes. The reaction solution was distilled off under reduced pressure,and the residue was neutralized with saturated aqueous NaHCO₃ solution,extracted with chloroform and the extract was washed with saturatedaqueous sodium chloride solution. The organic layer was dried overmagnesium sulfate, and then the solvent was distilled off under reducedpressure to give 3.51 g (96%) of N-Me-Lys (Z)-Tyr(3-tert-Bu)-N-Me-β-Ala-OBzl.

[0194]¹H-NMR (CDCl₃): δ 0.90-1.04 (2H, m), 1.20-1.60 (4H, m), 1.38 (9H,s), 2.30 (3H, s), 2.54-3.16 (7H, m), 2.81 and 2.89 (total 3H, s),3.50-3.80 (2H, m), 4.89 (1H, brs), 5.04-5.20 (1H, m), 5.11 and 5.13(total 4H, s), 6.60-7.08 (4H, m), 7.28-7.40 (10H, m).

[0195] (5) Synthesis of Boc-Phe (4-F)-N-Me-Lys (Z)-Tyr(3-tert-Bu)-N-Me-β-Ala-OBzl

[0196] To a solution of 3.40 g (4.94 mmol) of N-Me-Lys (Z)-Tyr(3-tert-Bu)-N-Me-β-Ala-OBzl and 1.47 g (5.19 mmol) of Boc-Phe (4-F)-OHin 25 ml of THF were added 1.39 g (5.43 mmol) of CMPI and 1.51 ml (10.9mmol) of TEA with ice cooling, and the mixture was stirred at roomtemperature for 2 hours. The reaction solution was combined with water,extracted with ethyl acetate and the extract was washed with saturatedaqueous sodium chloride solution. The organic layer was dried overmagnesium sulfate, and then the solvent was distilled off under reducedpressure, and the residue was purified by silica gel columnchromatography (developing solvent; ethyl acetate:hexane=1:1) to give4.22 g (90%) of Boc-Phe (4-F)-N-Me-Lys (Z)-Tyr(3-tert-Bu)-N-Me-β-Ala-OBzl.

[0197]¹H-NMR (CDCl₃): δ 0.80-1.90 (6H, m), 1.35 and 1.37 (total 18H, s),2.36-3.66 (16H, m), 4.60-5.40 (5H, m), 5.08 and 5.11 (total 4H, s),6.30-7.22 (7H, m), 7.30-7.42 (10H, m).

[0198] (6) Synthesis of(2S-(2S,13S))-2-tert-butoxycarbonylamino-N-(2-(3-tert-butyl-4-hydroxyphenylmethyl)-4-methyl-1,4,8-triaza-3,7,14-trioxocyclotetradec-13-yl)-3-(4-fluorophenyl)-N-methylpropionamide

[0199] To a solution of 4.00 g (4.19 mmol) of Boc-Phe (4-F)-N-Me-Lys(Z)-Tyr (3-tert-Bu)-N-Me-β-Ala-OBzl in 84 ml of methanol was added 400mg of 10% palladium on carbon, and the mixture was stirred overnight atroom temperature under a hydrogen atmosphere. After filtration, thefiltrate was concentrated under reduced pressure to give 3.05 g (100%)of Boc-Phe (4-F)-N-Me-Lys-Tyr (3-tert-Bu)-N-Me-β-Ala-OH.

[0200] To a mixed solution of 2.25 g (3.09 mmol) of the thus obtainedBoc-Phe (4-F)-N-Me-Lys-Tyr (3-tert-Bu)-N-Me-β-Ala-OH in 150 ml of DMFand 150 ml of pyridine was added 4.10 g (9.27 mmol) of BOP reagent, andthe mixture was stirred at room temperature for 16 hours. The reactionsolution was combined with water and extracted with ethyl acetate, andthen the extract was successively washed with 10% aqueous CUSO₄solution, saturated aqueous NaHCO₃ solution and saturated aqueous sodiumchloride solution. The organic layer was dried over magnesium sulfate,and then the solvent was distilled off under reduced pressure, and theresidue was purified by silica gel column chromatography (developingsolvent; chloroform:methanol=20:1) to give 1.32 g (60%) of(2S-(2S,13S))-2-tert-butoxycarbonylamino-N-(2-(3-tert-butyl-4-hydroxyphenylmethyl)-4-methyl-1,4,8-triaza-3,7,14-trioxocyclotetradec-13-yl)-3-(4-fluorophenyl)-N-methylpropionamide.

[0201]¹H-NMR (CDCl₃): δ 1.20-2.02 (6H, m), 1.30, 1.33, 1.37, 1.38 and1.47 (total 18H, s), 2.40-3.30 (16H, m), 3.60-3.78 (1H, m), 4.30-5.50(4H, m), 6.50-7.40 (7H, m).

[0202] (7) Synthesis of(2S-(2S,13S))-2-amino-N-(2-(3-tert-butyl-4-hydroxyphenylmethyl)-4-methyl-1,4,8-triaza-3,7,14-trioxocyclotetradec-13-yl)-3-(4-fluorophenyl)-N-methylpropionamide

[0203] To a solution of 1.20 g (1.69 mmol) of(2S-(2S,13S))-2-tert-butoxycarbonylamino-N-(2-(3-tert-butyl-4-hydroxyphenylmethyl)-4-methyl-1,4,8-triaza-3,7,14-trioxocyclotetradec-13-yl)-3-(4-fluorophenyl)-N-methylpropionamidein 15 ml of methylene chloride was added 5 ml of TFA, and the mixturewas stirred at room temperature for 40 minutes. The reaction solutionwas distilled off under reduced pressure, and the residue wasneutralized with saturated aqueous NaHCO₃ solution, extracted withchloroform and the extract was washed with saturated aqueous sodiumchloride solution. The organic layer was dried over magnesium sulfate,and then the solvent was distilled off under reduced pressure, and theresidue was purified by silica gel column chromatography (developingsolvent; chloroform:methanol:aqueous ammonia=20:1:0.1) to give 1.08 g(92%) of the title compound.

[0204] FAB-MS (M⁺+1): 612.

[0205]¹H-NMR (DMSO-d): δ 0.50-3.40 (16H, m), 1.29 (9H, s), 2.74, 2.78and 2.87 (total 6H, s), 3.77 and 4.42 (total 1H, t, J=6.5 Hz), 4.02-4.16(1H, m), 4.88-5.04 (1H, m), 6.61 (1H, d, J=8.0 Hz), 6.78 (1H, dd, J=2.0,8.0 Hz), 6.90-7.24 (4H, m), 7.40-7.56 (1H, m), 8.31 and 9.07 (total 1H,s), 8.40 and 8.79 (total 1H, d, J=8.9-9.2 Hz).

Example 5 Synthesis of(2S-(2S,13S))-2-amino-N-(2-(3-tert-butyl-4-hydroxyphenylmethyl)-8-methyl-1,4,8-triaza-3,7,14-trioxocyclotetradec-13-yl)-3-(4-fluorophenyl)-N-methylpropionamide

[0206] (1) Synthesis of Boc-N-Me-Lys (Me, Z)-OH

[0207] To a solution of 3.50 g (9.21 mmol) of Boc-Lys (Z)-OH in 86 ml ofTHF were added 1.37 g (34.3 mmol) of 60% sodium hydride and 4.27 ml(68.6 mmol) of methyl iodide with ice cooling, and the mixture wasstirred at room temperature for 15 hours. The reaction solution wascombined with 10% aqueous citric acid solution with ice cooling,extracted with ethyl acetate and the extract was washed with saturatedaqueous sodium chloride solution. The organic layer was dried overmagnesium sulfate, and then the solvent was distilled off under reducedpressure, and the residue was purified by silica gel columnchromatography (developing solvent; ethyl acetate:n-hexane:=1:1) to give3.60 g (96%) of Boc-N-Me-Lys (Me, Z)-OH.

[0208]¹H-NMR (CDCl₃): δ 1.20-2.02 (6H, m), 1.46 (9H, s), 2.79 (3H, s),2.91 (3H, s), 3.22-3.34 (2H, m), 4.34-4.70 (1H, m), 5.12 (2H, s),7.30-7.42 (5H, m).

[0209] (2) Synthesis of Boc-N-Me-Lys (Me, Z)-Tyr (3-tert-Bu)-β-Ala-OBzl

[0210] To a solution of 2.50 g (6.28 mmol) of H-Tyr(3-tert-Bu)-P-Ala-OBzl and 2.82 g (6.91 mmol) of Boc-N-Me-Lys (Me, Z)-OHin 18.8 ml of DMF were added 849 mg (6.28 mmol) of HOBt, 0.718 ml (6.28mmol) of NMM and 1.32 g (6.91 mmol) of WSCI with ice cooling, and themixture was stirred at room temperature for 90 minutes. The reactionsolution was combined with water, extracted with ethyl acetate and theextract was washed with 10% aqueous citric acid solution, saturatedaqueous NaHCO₃ solution and saturated aqueous sodium chloride solution.The organic layer was dried over magnesium sulfate, and then the solventwas distilled off under reduced pressure to give 4.81 g (97%) ofBoc-N-Me-Lys (Me, Z)-Tyr (3-tert-Bu)-β-Ala-OBzl.

[0211]¹H-NMR (CDCl₃): δ 1.10-1.90 (6H, m), 1.37 (9H, s), 1.45 (9H, s),2.40-2.60 (5H, m), 2.80-3.00 (2H, m), 2.88 and 2.96 (total 3H, s),3.16-3.50 (4H, m), 4.30-4.58 (2H, m), 5.09 and 5.11 (total 4H, s),6.30-6.68 (3H, m), 6.83 (1H, dd, J=1.7, 7.9 Hz), 7.03 (1H, d, J=1.7 Hz),7.30-7.42 (10H, m).

[0212] (3) Synthesis of N-Me-Lys (Me, Z)-Tyr (3-tert-Bu)-β-Ala-OBzl

[0213] To a solution of 4.48 g (5.69 mmol) of Boc-N-Me-Lys (Me, Z)-Tyr(3-tert-Bu)-β-Ala-OBzl in 30 ml of methylene chloride was added 10 ml ofTFA, and the mixture was stirred at room temperature for 40 minutes. Thereaction solution was distilled off under reduced pressure, and theresidue was neutralized with saturated aqueous NaHCO₃ solution,extracted with chloroform and the extract was washed with saturatedaqueous sodium chloride solution. The organic layer was dried overmagnesium sulfate, and then the solvent was distilled off under reducedpressure to give 3.80 g (97%) of N-Me-Lys (Me, Z)-Tyr(3-tert-Bu)-β-Ala-OBzl.

[0214]¹H-NMR (CDCl₃): δ 0.70-1.60 (6H, m), 1.38 (9H, s), 2.29 (3H, s),2.50-2.62 (2H, m), 2.68-2.90 (3H, m), 2.88 and 2.96 (total 3H, s),3.14-3.60 (4H, m), 4.40-4.70 (1H, m), 5.12 and 5.18 (total 4H, s),6.60-6.90 (3H, m), 7.02 (1H, s), 7.30-7.42 (10H, m).

[0215] (4) Synthesis of Boc-Phe (4-F)-N-Me-Lys (Me, Z)-Tyr(3-tert-Bu)-β-Ala-OBzl

[0216] To a solution of 3.51 g (5.10 mmol) of N-Me-Lys (Me, Z)-Tyr(3-tert-Bu)-β-Ala-OBzl and 1.52 g (5.36 mmol) of Boc-Phe (4-F)-OH in25.5 ml of THF were added 1.43 g (5.61 mmol) of CMPI and 1.56 ml (11.2mmol) of TEA with ice cooling, and the mixture was stirred at roomtemperature for 90 minutes. The reaction solution was combined withwater and extracted with ethyl acetate, and then the extract was washedwith saturated aqueous sodium chloride solution. The organic layer wasdried over magnesium sulfate, and then the solvent was distilled offunder reduced pressure, and the residue was purified by silica gelcolumn chromatography (developing solvent; ethyl acetate:hexane=1:1) togive 4.59 g (94%) of Boc-Phe (4-F)-N-Me-Lys (Me, Z)-Tyr(3-tert-Bu)-β-Ala-OBzl.

[0217]¹H-NMR (CDCl₃): δ 0.90-1.40 (5H, m), 1.30, 1.36 and 1.39 (total18H, s), 1.76-1.92 (1H, m), 2.10-3.60 (16H, m), 4.40-4.80 (2H, m), 4.99,5.11 and 5.13 (total 4H, s), 5.00-5.60 (2H, m), 6.50-7.40 (18H, m).

[0218] (5) Synthesis of(2S-(2S,13S))-2-(tert-butoxycarbonylamino)-N-(2-(3-tert-butyl-4-hydroxyphenylmethyl)-8-methyl-1,4,8-triaza-3,7,14-trioxocyclotetradec-13-yl)-3-(4-fluorophenyl)-N-methylpropionamide

[0219] To a solution of 4.32 g (4.53 mmol) of Boc-Phe (4-F)-pN-Me-Lys(Me, Z)-Tyr (3-tert-Bu)-β-Ala-OBzl in 50 ml of methanol was added 400 mgof 10% palladium on carbon, and the mixture was stirred at roomtemperature for 4 hours under a hydrogen atmosphere. After filtration,the filtrate was concentrated under reduced pressure to give 3.32 g(100%) of Boc-Phe (4-F)-N-Me-Lys (Me)-Tyr (3-tert-Bu)-β-Ala-OH.

[0220] To a mixed solution of 2.30 g (3.16 mmol) of the thus obtainedBoc-Phe (4-F)-N-Me-Lys (Me)-Tyr (3-tert-Bu)-β-Ala-OH in 150 ml of DMFand 150 ml of pyridine was added 4.19 g (9.48 mmol) of BOP reagent, andthe mixture was stirred at room temperature for 16 hours. The reactionsolution was combined with water, extracted with ethyl acetate and theextract was washed with saturated aqueous sodium chloride solution. Theorganic layer was dried over magnesium sulfate, and then the solvent wasdistilled off under reduced pressure, and the residue was purified bysilica gel column chromatography (developing solvent;chloroform:methanol=20:1) to give 811 mg (36%) of(2S-(2S,13S))-2-(tert-butoxycarbonylamino)-N-(2-(3-tert-butyl-4-hydroxyphenylmethyl)-8-methyl-1,4,8-triaza-3,7,14-trioxocyclotetradec-13-yl)-3-(4-fluorophenyl)-N-methylpropionamide.

[0221]¹H-NMR (CDCl₃): δ 0.80-2.00 (6H, m), 1.34, 1.37 and 1.40 (total18H, s), 2.30-3.40 (16H, m), 4.40-4.70 (2H, m), 5.16-5.44 (2H, m),6.40-7.20 (9H, m).

[0222] (6) Synthesis of(2S-(2S,13S))-2-amino-N-(2-(3-tert-butyl-4-hydroxyphenylmethyl)-8-methyl-1,4,8-triaza-3,7,14-trioxocyclotetradec-13-yl)-3-(4-fluorophenyl)-N-methylpropionamide

[0223] To a solution of 748 mg (1.05 mmol) of(2S-(2S,13S))-2-(tert-butoxycarbonylamino)-N-(2-(3-tert-butyl-4-hydroxyphenylmethyl)-8-methyl-1,4,8-triaza-3,7,14-trioxocyclotetradec-13-yl)-3-(4-fluorophenyl)-N-methylpropionamidein 10 ml of methylene chloride was added 5 ml of TFA, and the mixturewas stirred at room temperature for 40 minutes. The reaction solutionwas distilled off under reduced pressure, and the residue wasneutralized with saturated aqueous NaHCO₃ solution, extracted withchloroform and the extract was washed with saturated aqueous sodiumchloride solution. The organic layer was dried over magnesium sulfate,and then the solvent was distilled off under reduced pressure, and theresidue was purified by silica gel column chromatography (developingsolvent; chloroform:methanol=20:1) to give 608 mg (95%) of the titlecompound.

[0224] FAB-MS (M⁺+1): 612.

[0225]¹H-NMR (DMSO-d): δ 0.60-1.90 (6H, m), 1.26 and 1.28 (total 9H, s),2.20-2.90 (12H, m), 3.20-4.40 (6H, m), 4.80-4.90 (1H, m), 6.61 (1H, d,J=7.9 Hz), 6.70-7.60 (7H, m), 8.08-8.60 (1H, m), 9.01 and 9.11 (total1H, s).

Example 6 Synthesis of(2S-(2S,14S))-2-amino-N-(2-(3-tert-butyl-4-hydroxyphenylmethyl)-1,4,9-triaza-3,8,15-trioxocyclopentadec-14-yl)-3-(4-fluorophenyl)-N-methylpropionamide

[0226] (1) Synthesis of N-(Boc-3-tert-Bu-tyrosyl)-4-aminobutyric AcidBenzyl Ester

[0227] To a solution of 2.01 g (5.96 mmol) of Boc-Tyr (3-tert-Bu)-OH and1.51 g (6.56 mmol) of 4-aminobutyric acid benzyl ester hydrochloride(Helv. Chim. Acta., 80, 1997, 1253) in 24 ml of DMF were added 805 mg(5.96 mmol) of HOBt, 0.750 ml (6.56 mmol) of NMM and 1.26 g (6.57 mmol)of WSCI with ice cooling, and the mixture was stirred at roomtemperature for 90 minutes. The reaction solution was combined withwater and extracted with ethyl acetate, and then the extract wassuccessively washed with 10% aqueous citric acid solution, saturatedaqueous NaHCO₃ solution and saturated aqueous sodium chloride solution.The organic layer was dried over magnesium sulfate, and then the solventwas distilled off under reduced pressure to give 2.49 g (82%) ofN-(Boc-3-tert-Bu-tyrosyl)-4-aminobutyric acid benzyl ester.

[0228]¹H-NMR (CDCl₃): δ 1.37 (9H, s), 1.41 (9H, s), 1.62-1.78 (2H, m),2.26 (2H, t, J=7.3 Hz), 2.82-3.30 (4H, m), 4.20 (1H, q, J=7.3 Hz), 4.97(1H, brs), 5.10 (2H, s), 5.83 (1H, brs), 6.54 (1H, d, J=7.9 Hz), 6.86(1H, dd, J=1.7, 7.9 Hz), 7.04 (1H, d, J=1.7 Hz), 7.30-7.42 (5H, m).

[0229] (2) Synthesis of N-(3-tert-Bu-tyrosyl)-4-aminobutyric Acid BenzylEster

[0230] To a solution of 2.42 g (4.73 mmol) ofN-(Boc-3-tert-Bu-tyrosyl)-4-aminobutyric acid benzyl ester in 20 ml ofmethylene chloride was added 5 ml of TFA, and the mixture was stirred atroom temperature for 1 hour. The reaction solution was distilled offunder reduced pressure, and the residue was neutralized with saturatedaqueous NaHCO₃ solution, extracted with chloroform and the extract waswashed with saturated aqueous sodium chloride solution. The organiclayer was dried over magnesium sulfate, and then the solvent wasdistilled off under reduced pressure to give 1.94 g (100%) ofN-(3-tert-Bu-tyrosyl)-4-aminobutyric acid benzyl ester.

[0231]¹H-NMR (CDCl₃): δ 1.39 (9H, s), 1.85 (2H, tt, J=6.6, 7.6 Hz), 2.38(2H, t, J=7.6 Hz), 2.60 (1H, dd, J=9.2, 13.8 Hz), 3.14 (1H, dd, J=3.9,13.8 Hz), 3.30 (2H, q, J=6.6 Hz), 3.53 (1H, dd, J=3.9, 9.2 Hz), 5.12(2H, s), 6.61 (1H, d, J=7.9 Hz), 6.88 (1H, dd, J=2.0, 7.9 Hz), 7.07 (1H,d, J=2.0 Hz), 7.30-7.42 (5H, m).

[0232] (3) Synthesis ofN-(Boc-N-Me-Nε-Z-lysyl-3-tert-Bu-tyrosyl)-4-aminobutyric Acid BenzylEster

[0233] To a solution of 1.89 g (4.59 mmol) ofN-(3-tert-Bu-tyrosyl)-4-aminobutyric acid benzyl ester and 2.78 g (4.82mmol) of Boc-N-Me-Lys (Z)-OH in 14 ml of DMF were added 620 mg (4.59mmol) of HOBt, 0.787 ml (6.89 mmol) of NMM and 968 mg (5.05 mmol) ofWSCI with ice cooling, and the mixture was stirred at room temperaturefor 90 minutes. The reaction solution was combined with water andextracted with ethyl acetate, and then the extract was successivelywashed with 10% aqueous citric acid solution, saturated aqueous NaHCO₃solution and saturated aqueous sodium chloride solution. The organiclayer was dried over magnesium sulfate, and then the solvent wasdistilled off under reduced pressure to give 3.41 g (90%) ofN-(Boc-N-Me-Ne-Z-lysyl-3-tert-Bu-tyrosyl)-4-aminobutyric acid benzylester.

[0234]¹H-NMR (CDCl₃): δ 1.12-1.90 (8H, m), 1.36 (9H, s), 1.44 (9H, s),2.27 (2H, t, J=7.3 Hz), 2.56 (3H, s), 2.90-3.30 (6H, m), 4.37 (1H, t,J=7.1 Hz), 4.69 (1H, q, J=7.6 Hz), 4.82 (1H, brs), 5.09 (4H, s),5.50-5.70 and 6.00-6.20 (total 1H, m), 6.47 (1H, d, J=4.3 Hz), 6.59 (1H,d, J=7.9 Hz), 6.84 (1H, dd, J=1.7, 7.9 Hz), 7.04 (1H, d, J=1.7 Hz),7.30-7.44 (10H, m).

[0235] (4) Synthesis ofN-(N-Me-Ne-Z-lysyl-3-tert-Bu-tyrosyl)-4-aminobutyric Acid Benzyl Ester

[0236] To a solution of 3.32 g (4.21 mmol) ofN-(Boc-N-Me-Nε-Z-lysyl-3-tert-Bu-tyrosyl)-4-aminobutyric acid benzylester in 20 ml of methylene chloride was added 10 ml of TFA, and themixture was stirred at room temperature for 40 minutes. The reactionsolution was distilled off under reduced pressure, and the residue wasneutralized with saturated aqueous NaHCO₃ solution, extracted withchloroform and the extract was washed with saturated aqueous sodiumchloride solution. The organic layer was dried over magnesium sulfate,and then the solvent was distilled off under reduced pressure to give2.86 g (99%) of N-(N-Me-Nε-Z-lysyl-3-tert-Bu-tyrosyl)-4-aminobutyricacid benzyl ester.

[0237]¹H-NMR (CDCl₃): δ 0.80-0.98 (2H, m), 1.10-1.70 (4H, m), 1.38 (9H,s), 1.81 (2H, tt, J=6.6, 7.3 Hz), 2.28 (3H, s), 2.35 (2H, t, J=7.3 Hz),2.76-3.20 (5H, m), 3.27 (2H, q, J=6.6 Hz), 4.46-4.58 (1H, m), 4.80-4.94(1H, m), 5.11 and 5.14 (total 4H, s), 6.40-6.52 (1H, m), 6.69 (1H, d,J=7.9 Hz), 6.84 (1H, dd, J=1.6, 7.9 Hz), 6.88 (1H, brs), 7.04 (1H, d,J=1.6 Hz), 7.30-7.46 (10H, m).

[0238] (5) Synthesis ofN-(Boc-4-fluorophenylalanyl-N-Me-Nε-Z-lysyl-3-tert-Bu-tyrosyl)-4-aminobutyricAcid Benzyl Ester

[0239] To a solution of 2.82 g (4.10 mmol) ofN-(N-Me-Nε-Z-lysyl-3-tert-Bu-tyrosyl)-4-aminobutyric acid benzyl esterand 1.28 g (4.51 mmol) of Boc-Phe (4-F)-OH in 21 ml of THF were added1.26 g (4.92 mmol) of CMPI and 1.14 ml (8.20 mmol) of TEA with icecooling, and the mixture was stired at room temperature for 2 hours. Thereaction solution was combined with water and extracted with ethylacetate, and then the extract was washed with saturated aqueous sodiumchloride solution. The organic layer was dried over magnesium sulfate,and then the solvent was distilled off under reduced pressure, and theresidue was purified by silica gel column chromatography (developingsolvent; ethyl acetate:hexane=1:1) to give 3.57 g (91%) ofN-(Boc-4-fluorophenylalanyl-N-Me-Nε-Z-lysyl-3-tert-Bu-tyrosyl)-4-aminobutyricacid benzyl ester.

[0240]¹H-NMR (CDCl₃): δ 0.80-1.50 (6H, m), 1.35, 1.37 and 1.38 (total18H, s), 1.70-1.86 (2H, m), 2.15 and 2.62 (total 3H, s), 2.24-2.38 (2H,m), 2.60-3.30 (8H, m), 4.28-4.94 (3H, m), 5.26-5.40 (1H, m), 6.08-7.20(10H, m), 7.30-7.42 (10H, m).

[0241] (6) Synthesis of(2S-(2S,14S))-2-(tert-butoxycarbonylamino)-N-(2-(3-tert-butyl-4-hydroxyphenylmethyl)-1,4,9-triaza-3,8,15-trioxocyclopentadec-14-yl)-3-(4-fluorophenyl)-N-methylpropionamide

[0242] To a solution of 3.50 g (3.67 mmol) ofN-(Boc-4-fluorophenylalanyl-N-Me-Nε-Z-lysyl-3-tert-Bu-tyrosyl)-4-aminobutyricacid benzyl ester in 100 ml of methanol was added 400 mg of 10%palladium on carbon, and the mixture was stirred overnight at roomtemperature under a hydrogen atmosphere. After filtration, the filtratewas concentrated under reduced pressure to give 2.65 g (99%) ofN-(Boc-4-fluorophenylalanyl-N-Me-lysyl-3-tert-Bu-tyrosyl)-4-aminobutyricacid.

[0243] To a mixed solution of 2.35 g (3.22 mmol) of the thus obtainedN-(Boc-4-fluorophenylalanyl-N-Me-lysyl-3-tert-Bu-tyrosyl)-4-aminobutyricacid in 160 ml of DMF and 160 ml of pyridine was added 4.27 g (9.66mmol) of BOP reagent, and the mixture was stirred at room temperaturefor 16 hours. The reaction solution was combined with water andextracted with ethyl acetate, and then the extract was successivelywashed with 10% aqueous citric acid solution, saturated aqueous NaHCO₃solution and saturated aqueous sodium chloride solution. The organiclayer was dried over magnesium sulfate, and then the solvent wasdistilled off under reduced pressure, and the residue was purified bysilica gel column chromatography (developing solvent;chloroform:methanol=20:1) to give 1.88 g (82%) of(2S-(2S,14S))-2-(tert-butoxycarbonylamino)-N-(2-(3-tert-butyl-4-hydroxyphenylmethyl)-1,4,9-triaza-3,8,15-trioxocyclopentadec-14-yl)-3-(4-fluorophenyl)-N-methylpropionamide.

[0244]¹H-NMR (CDCl₃): δ 1.04-1.50 (6H, m), 1.29, 1.34 and 1.39 (total18H, s), 1.70-1.94 (2H, m), 2.10-2.40 (2H, m), 2.24 and 2.55 (total 3H,s), 2.56-3.70 (8H, m), 4.40-4.94 (3H, m), 5.24-5.40 (1H, m), 5.80-6.44(2H, m), 6.62-7.20 (7H, m), 8.21 (1H, d, J=8.2 Hz).

[0245] (7) Synthesis of(2S-(2S,14S))-2-amino-N-(2-(3-tert-butyl-4-hydroxyphenylmethyl)-1,4,9-triaza-3,8,15-trioxocyclopentadec-14-yl)-3-(4-fluorophenyl)-N-methylpropionamide

[0246] To a solution of 1.75 g (2.46 mmol) of(2S-(2S,14S))-2-(tert-butoxycarbonylamino)-N-(2-(3-tert-butyl-4-hydroxyphenylmethyl)-1,4,9-triaza-3,8,15-trioxocyclopentadec-14-yl)-3-(4-fluorophenyl)-N-methylpropionamidein 20 ml of methylene chloride was added 10 ml of TFA, and the mixturewas stirred at room temperature for 40 minutes. The reaction solutionwas distilled off under reduced pressure, and the residue wasneutralized with saturated aqueous NaHCO₃ solution, extracted withchloroform and the extract was washed with saturated aqueous sodiumchloride solution. The organic layer was dried over magnesium sulfate,and then the solvent was distilled off under reduced pressure, and theresidue was purified by silica gel column chromatography (developingsolvent; chloroform:methanol:aqueous ammonia=20:1:0.1) to give 783 mg(52%) of the title compound.

[0247] FAB-MS (M⁺+1): 612.

[0248]¹H-NMR (DMSO-d): δ 0.60-2.00 (6H, m), 1.06 (2H, t, J=6.9 Hz), 1.28and 1.29 (total 9H, s), 2.06-2.22 (2H, m), 2.50-3.00 (9H, m), 3.20-4.04(3H, m), 4.24-4.44 (3/2H, m), 4.96 (1/2H, d, J=9.9 Hz), 6.61 (1H, d,J=8.3 Hz), 6.70-6.84 (1H, m); 6.90-7.74 (7H, m), 8.14 and 8.95 (total1H, d, J=7.6-8.2 Hz), 9.04 and 9.09 (total 1H, s).

Example 7 Synthesis of(2S,14S)-13-(2S-2-amino-3-(4-fluorophenyl)propylamino)-2-(3-tert-butyl-4-hydroxyphenylmethyl)-1,4,8-triaza-3,7,14-trioxocyclotetradecane

[0249] (1) Synthesis of Z-Tyr (3-tert-Bu)-β-Ala-OtBu

[0250] To a solution of 2.69 g (7.25 mmol) of Z-Tyr (3-tert-Bu)-OH and1.45 g (7.98 mmol) of H-β-Ala-OtBu hydrochloride in 22 ml of DMF wereadded 980 mg (7.25 mmol) of HOBt, 1.66 ml (14.5 mmol) of NMM and 1.53 g(7.98 mmol) of WSCI with ice cooling, and the mixture was stirred atroom temperature for 90 minutes. The reaction solution was combined withwater and extracted with ethyl acetate, and then the extract wassuccessively washed with 10% aqueous citric acid solution, saturatedaqueous NaHCO₃ solution and saturated aqueous sodium chloride solution.The organic layer was dried over magnesium sulfate, and then the solventwas distilled off under reduced pressure to give 2.76 g (76%) of Z-Tyr(3-tert-Bu)-β-Ala-OtBu.

[0251]¹H-NMR (CDCl₃): δ 1.37 (9H, s), 1.41 (9H, s), 2.33 (2H, q, J=6.3Hz), 2.88-3.06 (2H, m), 3.40 (2H, q, J=6.3 Hz), 4.20-4.34 (1H, m),4.80-5.00 (1H, m), 5.09. (2H, s), 5.20-5.30 (1H, m), 6.12-6.24 (1H, m),6.57 (1H, d, J=7.9 Hz), 6.86 (1H, d, J=7.9 Hz), 7.03 (1H, s), 7.30-7.42(5H, m).

[0252] (2) Synthesis of H-Tyr (3-tert-Bu)-β-Ala-OtBu

[0253] To a solution of 2.70 g (5.42 mmol) of Z-Tyr(3-tert-Bu)-β-Ala-OtBu in 80 ml of methanol was added 300 mg of 10%palladium on carbon, and the mixture was stirred at room temperature for5 hours under a hydrogen atmosphere. After filtration, the filtrate wasconcentrated under reduced pressure to give 1.96 g (99%) of H-Tyr(3-tert-Bu)-p-Ala-OtBu.

[0254]¹H-NMR (CDCl₃): δ 1.36 (9H, s), 1.43 (9H, s), 2.40 (2H, t, J=6.6Hz), 2.77 (1H, dd, J=8.2, 13.5 Hz), 3.14 (1H, dd, J=4.6, 13.5 Hz),3.36-3.50 (2H, m), 3.76-3.84 (1H, m), 6.71 (1H, d, J=7.9 Hz), 6.93 (1H,d, J=7.9 Hz), 7.04 (1H, s), 7.64-7.72 (1H, m).

[0255] (3) Synthesis of Z-N-Me-Lys (Boc)-Tyr (3-tert-Bu)-β-Ala-OtBu

[0256] To a solution of 1.90 g (5.22 mmol) of H-Tyr(3-tert-Bu)-β-Ala-OtBu and 2.18 g (5.74 mmol) of Z-N-Me-Lys (Boc)-OH in17.5 ml of DMF were added 705 mg (5.22 mmol) of HOBt, 0.656 ml (5.74mmol) of NMM and 1.10 g (8.14 mmol) of WSCI with ice cooling, and themixture was stirred at room temperature for 1 hour. The reactionsolution was combined with water and extracted with ethyl acetate, andthen the extract was successively washed with 10% aqueous citric acidsolution, saturated aqueous NaHCO₃ solution and saturated aqueous sodiumchloride solution. The organic layer was dried over magnesium sulfate,and then the solvent was distilled off under reduced pressure to give3.49 g (92%) of Z-N-Me-Lys (Boc)-Tyr (3-tert-Bu)-β-Ala-OtBu.

[0257]¹H-NMR (CDCl₃): δ 0.90-1.10 (2H, m), 1.20-1.50 (4H, m), 1.37 (9H,s), 1.43 (9H, s), 1.44 (9H, s), 2.34-2.46 (2H, m), 2.80 (1H, dd, J=9.2,14.2 Hz), 2.99 (2H, q, J=6.6 Hz), 3.10-3.24 (1H, m), 3.43 (2H, q, J=6.0Hz), 3.94-4.04 (1H, m), 4.55 (1H, q, J=6.2 Hz), 4.65 (1H, brs), 5.10(2H, s), 5.20-5.30 (1H, m), 6.34 (1H, d, J=6.0 Hz), 6.42 (1H, brs), 6.60(1 h, brs), 6.66 (1H, d, J=7.9 Hz), 6.82 (1H, dd, J=1.7, 7.9 Hz), 7.01(1H, d, J=1.7 Hz), 7.30-7.42 (5H, m).

[0258] (4) Synthesis of(2S,14S)-13-amino-2-(3-tert-butyl-4-hydroxyphenylmethyl)-1,4,8-triaza-3,7,14-trioxocyclotetradecane

[0259] To a solution of 2.51 g (3.46 mmol) of Z-N-Me-Lys (Boc)-Tyr(3-tert-Bu)-β-Ala-OtBu in 10 ml of methylene chloride was added 10 ml ofTFA, and the mixture was stirred at room temperature for 50 minutes. Thereaction solution was distilled off under reduced pressure, and a mixedsolution of the residue in 170 ml of acetonitrile and 170 ml of pyridinewas stirred with 4.59 g (10.4 mmol) of BOP reagent at room temperaturefor 21 hours. The solvent was distilled off under reduced pressure, andthe residue was successively washed with diethyl ether and chloroform togive solids. The thus obtained solids were dissolved in 50 ml ofmethanol and combined with 300 mg of 10% palladium on carbon, and themixture was stirred overnight at room temperature under a hydrogenatmosphere. After filtration, the filtrate was concentrated underreduced pressure, and the residue was purified by silica gel columnchromatography (developing solvent; chloroform:methanol aqueousammonia=10:1:0.1) to give 1.30 g (90%) of(2S,14S)-13-amino-2-(3-tert-butyl-4-hydroxyphenylmethyl)-1,4,8-triaza-3,7,14-trioxocyclotetradecane.

[0260]¹H-NMR (CD₃OD): δ 0.90-1.40 (6H, m), 1.40 (9H, s), 2.28-2.50 (2H,m), 2.78 (1H, dd, J=7.6, 13.9 Hz), 3.06 (2H, dd, J=6.9, 13.5 Hz),3.30-3.72 (4H, m), 4.65 (1H, t, J=7.6 Hz), 6.66 (1H, d, J=7.9 Hz), 6.90(1H, dd, J=1.7, 7.9 Hz), 7.08 (1H, d, J=1.7 Hz).

[0261] (5) Synthesis of(2S,14S)-13-((2S)-2-(tert-butoxycarbonylamino)-3-(4-fluorophenyl)propylamino)-2-(3-tert-butyl-4-hydroxyphenylmethyl)-1,4,8-triaza-3,7,14-trioxocyclotetradecane

[0262] To a solution of 0.89 g (2.13 mmol) of(2S,14S)-13-amino-2-(3-tert-butyl-4-hydroxyphenylmethyl)-1,4,8-triaza-3,7,14-trioxocyclotetradecaneand 739 mg (2.77 mmol) of Boc-Phe (4-F)—H in 22 ml of methanol wereadded 1 ml of acetic acid and 423 mg (6.39 mmol) of 95% sodiumcyanoborohydride with ice cooling, and the mixture was stirred at roomtemperature for 30 minutes. The reaction solution was combined withwater and extracted with chloroform, and then the extract was washedwith saturated aqueous sodium chloride solution. The organic layer wasdried over magnesium sulfate, and then the solvent was distilled offunder reduced pressure, and the residue was purified by silica gelcolumn chromatography (developing solvent; chloroform:methanol:aqueousammonia=20:1:0.1) to give 793 mg (56%) of(2S,14S)-13-((2S)-2-(tert-butoxycarbonylamino)-3-(4-fluorophenyl)propylamino)-2-(3-tert-butyl-4-hydroxyphenylmethyl)-1,4,8-triaza-3,7,14-trioxocyclotetradecane.

[0263]¹H-NMR (CDCl₃): δ 1.12-1.80 (6H, m), 2.06 (1H, dd, J=5.3, 11.9Hz), 2.18 (1H, dd, J=6.3, 11.9 Hz), 2.28-2.64 (3H, m), 2.70-2.80 (2H,m), 2.94-3.12 (3H, m), 3.42-3.68 (3H, m), 4.72 (1H, dd, J=6.3, 9.3 Hz),6.65 (1H, d, J=7.9 Hz), 6.88 (1H, dd, J=2.0, 7.9 Hz), 7.00 (2 h, t,J=8.2 Hz), 7.11 (1H, d, J=2.0 Hz), 7.21 (2H, dd, J=7.6, 8.2 Hz).

[0264] (6) Synthesis of(2S,14S)-13-((2S)-2-amino-3-(4-fluorophenyl)propylamino)-2-(3-tert-butyl-4-hydroxyphenylmethyl)-1,4,8-triaza-3,7,14-trioxocyclotetradecane

[0265] To a solution of 735 mg (1.10 mmol) of(2S,14S)-13-((2S)-2-(tert-butoxycarbonylamino)-3-(4-fluorophenyl)propylamino)-2-(3-tert-butyl-4-hydroxyphenylmethyl)-1,4,8-triaza-3,7,14-trioxocyclotetradecanein 5 ml of methylene chloride was added 3 ml of TFA, and the mixture wasstirred at room temperature for 40 minutes. The reaction solution wasdistilled off under reduced pressure, and the residue was neutralizedwith saturated aqueous NaHCO₃ solution, extracted with chloroform andthe extract was washed with saturated aqueous sodium chloride solution.The organic layer was dried over magnesium sulfate, and then the solventwas distilled off under reduced pressure, and the residue was purifiedby silica gel column chromatography (developing solvent;chloroform:methanol:aqueous ammonia=5:1:0.1) to give 608 mg (97%) of thetitle compound.

[0266] FAB-MS (M⁺+1): 570.

[0267]¹H-NMR (DMSO-d): δ 1.00-1.52 (4H, m), 1.30 (9H, s), 1.64-1.94 (4H,m), 2.14-2.32 (3H, m), 2.50-2.90 (5H, m), 3.20-3.62 (4H, m), 4.42-4.56(1H, m), 6.63 (1H, d, J=8.2 Hz), 6.77 (1H, d, J=8.2 Hz), 6.93 (1H, s),7.00-7.24 (5H, m), 7.69 (1H, d, J=5.9 Hz), 8.09 (1H, d, J=9.2 Hz),9.00-9.20 (1H, m).

Example 8 Synthesis of(2S-(2S.13S))-N-(8-acetyl-2-(3-tert-butyl-4-hydroxyphenylmethyl)-N-methyl-1,4,8-triaza-3.14-dioxocyclotetradec-13-yl)-2-amino-3-(4-fluorophenyl)propionamide

[0268] (1) Synthesis of Boc-N-Me-Lys (Z)-Tyr (3-tert-Bu)-OMe

[0269] To a solution of 2.00 g (7.97 mmol) of H-Tyr (3-tert-Bu)-OMe and5.05 g (8.77 mmol) of Boc-N-Me-Lys (Z)-OH in 24 ml of DMF were added1.08 g (7.97 mmol) of HOBt, 1.37 ml (12.0 mmol) of NMM and 1.68 g (8.77mmol) of WSCI with ice cooling, and the mixture was stirred at roomtemperature for 3 hours. The reaction solution was combined with waterand extracted with ethyl acetate, and then the extract was successivelywashed with 10% aqueous citric acid solution, saturated aqueous NaHCO₃solution and saturated aqueous sodium chloride solution. The organiclayer was dried over magnesium sulfate, and then the solvent wasdistilled off under reduced pressure, and the residue was purified bysilica gel column chromatography (developing solvent; ethylacetate:hexane 1:1) to give 4.99 g (100%) of Boc-N-Me-Lys (Z)-Tyr(3-tert-Bu)-OMe.

[0270]¹H-NMR (CDCl₃): δ 1.14-1.30 (2H, m), 1.37 (9H, s), 1.43 (9H, s),1.40-1.64 (3H, m), 1.78-1.96 (1H, m), 2.56 (3H, s), 2.93 (1H, dd, J=7.3,14.2 Hz), 3.04-3.22 (3H, m), 3.73 (3H, s), 4.44 (1H, t, J=7.6 Hz),4.72-4.86 (2H, m), 5.09 (2H, s), 5.55 (1H, brs), 6.40 (1H, brs), 6.61(1H, d, J=7.9 Hz), 6.77 (1 h, d, J=7.9 Hz), 6.95 (1H, s), 7.30-7.40 (5H,m).

[0271] (2) Synthesis of Boc-N-Me-Lys-Tyr (3-tert-Bu)-OMe

[0272] To a solution of 4.92 g (7.85 mmol) of Boc-N-Me-Lys (Z)-Tyr(3-tert-Bu)-OMe in 80 ml of methanol was added 500 mg of 10% palladiumon carbon, and the mixture was stirred overnight at room temperatureunder a hydrogen atmosphere. After filtration, the filtrate wasconcentrated under reduced pressure to give 3.87 g (100%) ofBoc-N-Me-Lys-Tyr (3-tert-Bu)-OMe.

[0273]¹H-NMR (CDCl₃): δ 1.20-1.94 (6H, m), 1.36 (9H, s), 1.43 (9H, s),2.63 (3H, s), 2.80-3.00 (3H, m), 3.12 (1H, dd, J=5.0, 13.8 Hz), 3.74(3H, s), 4.45 (1H, t, J=6.9 Hz), 4.70-4.80 (1H, m), 6.73 (1H, d, J=7.9Hz), 6.82 (1H, d, J=7.9 Hz), 6.94 (1H, s).

[0274] (3) Synthesis ofBoc-N-Me-Nε-(3-benzyloxycarbonylaminopropyl)-Lys-Tyr (3-tert-Bu)-OMe

[0275] To a solution of 3.10 g (6.29 mmol) of Boc-N-Me-Lys-Tyr(3-tert-Bu)-OMe and 1.43 g (6.92 mmol) ofN-phenylmethoxycarbonyl-3-aminopropanal in 63 ml of methanol were added0.36 ml of acetic acid and 832 mg (12.6 mmol) of 95% sodiumcyanoborohydride with ice cooling, and the mixture was stirred at roomtemperature overnight. The reaction solution was combined with water andextracted with chloroform, and then the extract was washed withsaturated aqueous sodium chloride solution. The organic layer was driedover magnesium sulfate, and then the solvent was distilled off underreduced pressure, and the residue was purified by silica gel columnchromatography (developing solvent; chloroform:methanol:aqueousammonia=20:1:0.1) to give 2.30 g (53%) ofBoc-N-Me-Ne-(3-benzyloxycarbonylaminopropyl)-Lys-Tyr (3-tert-Bu)-OMe.

[0276]¹H-NMR (CDCl): δ 1.16-2.00 (8H, m), 1.37 (9H, s), 1.44 (9H, s),2.61 (3H, s), 2.72 (2H, t, J=7.3 Hz), 2.85 (2H, t, J=6.6 Hz), 2.84-2.94(1H, m), 3.12 (1H, dd, J=5.3, 13.9 Hz), 3.24-3.38 (2H, m), 3.74 (3H, s),4.42 (1H, t, J=7.3 Hz), 4.75 (1H, q, J=6.6 Hz), 5.10 (2H, s), 5.40-5.50(1H, m), 6.56-6.66 (1H, m), 6.70 (1H, d, J=7.9 Hz), 6.78 (1H, dd, J=1.7,7.9 Hz), 6.95 (1H, d, J=1.7 Hz), 7.28-7.40 (5H, m).

[0277] (4) Synthesis ofBoc-N-Me-Nε-(3-benzyloxycarbonylaminopropyl)-Ne-acetyl-Lys-Tyr(3-tert-Bu)-OMe

[0278] To a solution of 2.24 g (3.27 mmol) ofBoc-N-Me-Nε-(3-benzyloxycarbonylaminopropyl)-Lys-Tyr (3-tert-Bu)-OMe in33 ml of methylene chloride were added 0.401 ml (4.25 mmol) of aceticanhydride and 0.684 ml (4.91 mmol) of TEA with ice cooling, and themixture was stirred for 30 minutes. The reaction solution wasneutralized with saturated aqueous NaHCO₃ solution, extracted withmethylene chloride and the extract was washed with saturated aqueoussodium chloride solution. The organic layer was dried over magnesiumsulfate, and then the solvent was distilled off under reduced pressure,and the residue was purified by silica gel column chromatography(developing solvent; chloroform:methanol:aqueous ammonia=30:1:0.1) togive 1.92 g (81%) ofBoc-N-Me-Ne-acetyl-Nε-(3-benzyloxycarbonylaminopropyl)-Lys-Tyr(3-tert-Bu)-OMe.

[0279]¹H-NMR (CDCl₃): δ 1.08-2.00 (8H, m), 1.37 (9H, s), 1.43 (9H, s),2.05 (3.H, s), 2.96 (1H, dd, J=6.9, 14.5 Hz), 3.04-3.42 (7H, m), 3.73(3H, s), 4.30-4.60 (1H, m), 4.76 (1H, q, J=6.3 Hz), 5.09 (2H, s), 5.77(1H, brs), 6.30-6.50 (1H, m), 6.58-6.82 (2H, m), 6.95 (1H, s), 7.30-7.40(5H, m).

[0280] (5) Synthesis ofN-Me-Ne-acetyl-Nε-(3-benzyloxycarbonylaminopropyl)-Lys-Tyr(3-tert-Bu)-OMe

[0281] To a solution of 1.82 g (2.50 mmol) ofBoc-N-Me-Nε-(3-benzyloxycarbonylaminopropyl)-Nε-acetyl-Lys-Tyr(3-tert-Bu)-OMe in 20 ml of methylene chloride was added 10 ml of TFA,and the mixture was stirred at room temperature for 40 minutes. Thereaction solution was distilled off under reduced pressure, and theresidue was neutralized with saturated aqueous NaHCO₃ solution,extracted with chloroform and the extract was washed with saturatedaqueous sodium chloride solution. The organic layer was dried overmagnesium sulfate, and then the solvent was distilled off under reducedpressure to give 1.56 g (99%) ofN-Me-Nε-acetyl-Nε-(3-benzyloxycarbonylaminopropyl)-Lys-Tyr(3-tert-Bu)-OMe.

[0282]¹H-NMR (CDCl₃): δ 0.74-1.82 (8H, m), 1.38 and 1.39 (total 9H, s),2.02 and 2.07 (total 3H, s), 2.35 and 2.37 (total 3H, s), 2.60-3.40 (9H,m), 3.76 and 3.77 (total 3H, s), 4.78-5.00 (3/2H, m), 5.12 (2H, s),5.52-5.60 (1/2H, m), 6.60-7.04 (7/2H, m), 7.30-7.46 (5H, m), 7.67 (1/2H,m).

[0283] (6) Synthesis of Boc-Phe(4-F)-N-Me-Nε-acetyl-Nε-(3-benzyloxydarbonylaminopropyl)-Lys-Tyr(3-tert-Bu)-OMe

[0284] To a solution of 1.50 g (2.39 mmol) ofN-Me-Nε-acetyl-Nε-(3-benzyloxycarbonylaminopropyl)-Lys-Tyr(3-tert-Bu)-OMe and 811 mg (2.87 mmol) of Boc-Phe (4-F)-OH in 16 ml ofTHF were added 794 mg (3.11 mmol) of CMPI and 0.733 ml (5.26 mmol) ofTEA with ice cooling, and the mixture was stirred at room temperaturefor 14 hours. The reaction solution was combined with water andextracted with ethyl acetate, and then the extract was washed withsaturated aqueous sodium chloride solution. The organic layer was driedover magnesium sulfate, and then the solvent was distilled off underreduced pressure, and the residue was purified by silica gel columnchromatography (developing solvent; ethyl acetate:hexane=2:1) to give1.96 g (92%) of Boc-Phe(4-F)-N-Me-Nε-acetyl-Nε-(3-benzyloxycarbonylaminopropyl)-Lys-Tyr(3-tert-Bu)-OMe.

[0285]¹H-NMR (CDCl₃): δ 1.00-1.90 (8H, m), 1.35, 1.36 and 1.38 (total18H, s), 2.04 (3H, s), 2.20 and 2.55 (total 3H, s), 2.66-3.42 (10H, m),3.70 and 3.76 (total 3H, s), 4.60-5.04 (3H, m), 5.10 (2H, s), 5.20-6.24(3H, m), 6.58-7.20 (7H, m), 7.30-7.44 (5H, m).

[0286] (7) Synthesis of Boc-Phe(4-F)-N-Me-Nε-acetyl-Nε-(3-aminopropyl)-Lys-Tyr (3-tert-Bu)-OH

[0287] To a solution of 1.89 g (2.12 mmol) of Boc-Phe(4-F)-N-Me-Nε-acetyl-Nε-(3-benzyloxycarbonylaminopropyl)-Lys-Tyr(3-tert-Bu)-OMe in 20 ml of dioxane was added 2 ml of 2N aqueous sodiumhydroxide solution, and the mixture was stirred at room temperature for100 minutes. The reaction solution was combined with 10% aqueous citricacid solution with ice cooling, extracted with chloroform and theextract was washed with saturated aqueous sodium chloride solution. Theorganic layer was dried over magnesium sulfate, and then the solvent wasdistilled off under reduced pressure to give 1.87 g (100%) of Boc-Phe(4-F)-N-Me-Nε-acetyl-Nε-(3-benzyloxycarbonylaminopropyl)-Lys-Tyr(3-tert-Bu)-OH.

[0288] To a solution of 1.82 g (2.08 mmol) of Boc-Phe(4-F)-N-Me-Nε-acetyl-Nε-(3-benzyloxycarbonylaminopropyl)-Lys-Tyr(3-tert-Bu)-OH in 20 ml of methanol was added 200 mg of 10% palladium oncarbon, and the mixture was stirred overnight at room temperature undera hydrogen atmosphere. After filtration, the filtrate was concentratedunder reduced pressure to give 1.32 g (85%) of Boc-Phe(4-F)-N-Me-Nε-acetyl-Nε-(3-aminopropyl)-Lys-Tyr (3-tert-Bu)-OH.

[0289]¹H-NMR (DMSO-d): δ 1.00-2.00 (8H, m), 1.28 and 1.38 (total 18H,s), 1.96 (3H, s), 2.30-3.50 (9H, m), 4.00-5.00 (3H, m), 6.54-6.98 (3H,m), 7.00-7.30 (4H, m), 7.30-7.44 (1H, m), 8.96 (1H, d, J=5.3 Hz).

[0290] (8) Synthesis of(2S-(2S,13S))—N-(8-acetyl-2-(3-tert-butyl-4-hydroxyphenylmethyl)-3,14-dioxo-1,4,8-triazacyclotetradec-13-yl)-2-tert-butoxycarbonylamino-3-(4-fluorophenyl)-N-methylpropionamide

[0291] To a mixed solution of 1.10 g (1.48 mmol) of Boc-Phe(4-F)-N-Me-Nε-acetyl-Nε-(3-aminopropyl)-Lys-Tyr (3-tert-Bu)-OH in 75 mlof DMF and 75 ml of pyridine was added 1.96 g (4.44 mmol) of BOPreagent, and the mixture was stirred at room temperature for 18 hours.The reaction solution was concentrated under reduced pressure, combinedwith water and extracted with ethyl acetate, and then the extract wassuccessively washed with 10% aqueous citric acid solution and saturatedaqueous sodium chloride solution. The organic layer was dried overmagnesium sulfate, and then the solvent was distilled off under reducedpressure, and the residue was purified by silica gel columnchromatography (developing solvent; methylene chloride:methanol:aqueousammonia=20:1:0.05) to give 0.59 g (55%) of2S-(2S,13S))-N-(8-acetyl-2-(3-tert-butyl-4-hydroxyphenylmethyl)-3,14-dioxo-1,4,8-triazacyclotetradec-13-yl)-2-tert-butoxycarbonylamino-3-(4-fluorophenyl)-N-methylpropionamide.

[0292] (9) Synthesis of(2S-(2S,13S))-N-(8-acetyl-2-(3-tert-butyl-4-hydroxyphenylmethyl)-3,14-dioxo-1,4,8-triazacyclotetradec-13-yl)-2-amino-3-(4-fluorophenyl)-N-methylpropionamide

[0293] To a solution of 0.55 g (0.757 mmol) of(2S-(2S,13S))—N-(8-acetyl-2-(3-tert-butyl-4-hydroxyphenylmethyl)-3,14-dioxo-1,4,8-triazacyclotetradec-13-yl)-2-tert-butoxycarbonylamino-3-(4-fluorophenyl)-N-methylpropionamidein 7 ml of methylene chloride was added 3 ml of TFA, and the mixture wasstirred at room temperature for 55 minutes. The reaction solution wasdistilled off under reduced pressure, and the residue was neutralizedwith saturated aqueous NaHCO₃ solution, extracted with methylenechloride and the extract was washed with saturated aqueous sodiumchloride solution. The organic layer was dried over magnesium sulfate,and then the solvent was distilled off under reduced pressure, and theresidue was purified by silica gel column chromatography (developingsolvent; methylene chloride:methanol:aqueous ammonia=15:1:0.1) to give0.43 g (90%) of the title compound.

[0294] FAB-MS (M⁺+1): 626.

[0295]¹H-NMR (DMSO-d6—CDCl₃): δ 0.7-1.8 (16H, m), 1.29 and 1.30 (total9H, s), 1.9-2.0 (3H, m), 2.4-3.4 (10H, m), 2.62 and 2.64 (3H, s), 3.70(0.5H, m), 4.0-4.4 (2H, m), 4.92 (0.5H, d, J=9.2 Hz), 6.5-7.2 (8H, m),7.8-8.1 (1.5H, m), 9.0-9.1 (1.5H, m).

Example 9 Synthesis of(2S-(2S,13S))-2-amino-N-(2-(3-tert-butyl-4-hydroxyphenylmethyl)-9,N-dimethyl-1,4,9-triaza-3,8,14-trioxocyclotetradec-13-yl)-3-(4-fluorophenyl)propionamide

[0296] (1) Synthesis of Boc-N-Me-Orn (Me) (Z)-OH

[0297] To a solution of 3.00 g (8.19 mmol) of Boc-Orn (Z)-OH in 82 ml ofTHF were added 1.31 g (32.8 mmol) of 60% sodium hydride and 4.08 ml(65.5 mmol) of methyl iodide with ice cooling, and the mixture wasstirred at room temperature for 16 hours. The reaction solution wascombined with water with ice cooling, and washed with hexane. Theaqueous layer was combined with 10% aqueous citric acid solution,extracted with ethyl acetate and the extract was washed with saturatedaqueous sodium chloride solution. The organic layer was dried overmagnesium sulfate, and then the solvent was distilled off under reducedpressure to give 3.22 g (100%) of Boc-N-Me-Orn (Me) (Z)-OH.

[0298]¹H-NMR (CDCl₃): δ 1.46 (9H, s), 1.50-2.02 (4H, m), 2.70-2.90 (3H,m), 2.92 (3H, s), 3.20-3.46 (2H, m), 4.32-4.80 (1H, m), 5.13 (2H, s),7.24-7.40 (5H, m).

[0299] (2) Synthesis ofN-(Boc-N-Me-Ne-Z-ornithyl-3-tert-Bu-tyrosyl)-4-aminobutyric Acid BenzylEster

[0300] To a solution of 2.39 g (5.80 mmol) ofN-(3-tert-Bu-tyrosyl)-4-aminobutyric acid benzyl ester and 2.51 g (6.38mmol) of Boc-N-Me-Orn (Z)-OH in 29 ml of DMF were added 784 mg (5.80mmol) of HOBt, 0.729 ml (6.38 mmol) of NMM and 1.22 g (6.38 mmol) ofWSCI with ice cooling, and the mixture was stirred at room temperaturefor 140 minutes. The reaction solution was combined with water andextracted with ethyl acetate, and then the extract was successivelywashed with 10% aqueous citric acid solution, saturated aqueous NaHCO₃solution and saturated aqueous sodium chloride solution. The organiclayer was dried over magnesium sulfate, and then the solvent wasdistilled off under reduced pressure, and the residue was purified bysilica gel column chromatography (developing solvent; ethylacetate:hexane=1:1) to give 3.91 g (86%) ofN-(Boc-N-Me-Ne-Z-ornithyl-3-tert-Bu-tyrosyl)-4-aminobutyric acid benzylester.

[0301]¹H-NMR (CDCl₃): δ 1.36 (9H, s), 1.45 (9H, s), 1.50-1.90 (4H, m),2.26 (2H, t, J=7.3 Hz), 2.47 and 2.58 (total 3H, s), 2.88 (3H, s), 2.95(2H, d, J=7.3 Hz), 3.10-3.44 (4H, m), 4.42-4.60 (2H, m), 5.09 (2H, s),5.12 (2H, s), 5.30-5.50 (1H, m), 5.90-6.10 (1H, m), 6.54 (1H, d, J=7.9Hz), 6.83 (1H, d, J=7.9 Hz), 7.04 (1H, s), 7.24-7.40 (10H, m).

[0302] (3) Synthesis ofN-(N-Me-Nε-Z-ornithyl-3-tert-Bu-tyrosyl)-4-aminobutyric Acid BenzylEster

[0303] To a solution of 3.81 g (4.84 mmol) ofN-(Boc-N-Me-Nε-Z-ornithyl-3-tert-Bu-tyrosyl)-4-aminobutyric acid benzylester in 40 ml of methylene chloride was added 10 ml of TFA, and themixture was stirred at room temperature for 40 minutes. The reactionsolution was distilled off under reduced pressure, and the residue wasneutralized with saturated aqueous NaHCO₃ solution, extracted withchloroform and the extract was washed with saturated aqueous sodiumchloride solution. The organic layer was dried over magnesium sulfate,and then the solvent was distilled off under reduced pressure to give3.26 g (98%) of N-(N-Me-Nε-Z-ornithyl-3-tert-Bu-tyrosyl)-4-aminobutyricacid benzyl ester.

[0304]¹H-NMR (CDCl₃): δ 1.06-1.40 (2H, m), 1.37 (9H, s), 1.78 (2H, tt,J=6.9, 7.3 Hz), 2.27 (3H, s), 2.32 (2H, t, J=7.3 Hz), 2.76-2.92 (5H, m),2.98-3.30 (3H, m), 3.225 (2H, q, J=6.9 Hz), 4.34-4.56 (1H, m), 5.10 and5.14 (total 4H, s), 6.30-6.88 (3 h, m), 7.02 (1H, d, J=2.6 Hz),7.30-7.62 (11H, m).

[0305] (4) Synthesis ofN-(Boc-4-fluorophenylalanyl-N-Me-Nε-Z-ornithyl-3-tert-Bu-tyrosyl)-4-aminobutyricAcid Benzyl Ester

[0306] To a solution of 3.10 g (4.51 mmol) ofN-(N-Me-Nε-Z-ornithyl-3-tert-Bu-tyrosyl)-4-aminobutyric acid benzylester and 1.41 g (4.96 mmol) of Boc-Phe (4-F)-OH in 45 ml of THF wereadded 1.38 g (5.41 mmol) of CMPI and 1.38 ml (9.92 mmol) of TEA with icecooling, and the mixture was stirred at room temperature for 14 hours.The reaction solution was combined with water and extracted with ethylacetate, and then the extract was washed with saturated aqueous sodiumchloride solution. The organic layer was dried over magnesium sulfate,and then the solvent was distilled off under reduced pressure, and theresidue was purified by silica gel column chromatography (developigsolvent; ethyl acetate:hexane=2:1) to give 4.20 g (98%) ofN-(Boc-4-fluorophenylalanyl-N-Me-Nε-Z-ornithyl-3-tert-Bu-tyrosyl)-4-aminobutyricacid benzyl ester.

[0307]¹H-NMR (CDCl₃): δ 1.20-1.50 (2H, m), 1.34 and 1.38 (total 18H, s),1.60-1.84 (4H, m), 2.04-3.40 (16H, m), 4.30-5.06 (5/2H, m), 5.08 and5.11 (total 4H, s), 5.29 (1/2H, d, J=6.6 Hz), 5.60-6.30 (2H, m),6.40-7.20 (7H, m), 7.24-7.42 (21/2H, m), 7.68-7.76 (1/2H, m).

[0308] (5)(2S-(2S,13S))-2-tert-butoxycarbonylamino-N-(2-(3-tert-butyl-4-hydroxyphenylmethyl)-9-methyl-1,4,9-triaza-3,8,14-trioxocyclotetradec-13-yl)-3-(4-fluorophenyl)-N-methylpropionamide

[0309] To a solution of 2.80 g (2.68 mmol) ofN-(Boc-4-fluorophenylalanyl-N-Me-Nε-Z-ornithyl-3-tert-Bu-tyrosyl)-4-aminobutyricacid benzyl ester in 40 ml of methanol was added 0.52 g of palladiumhydroxide on carbon, and the mixture was stirred at room temperature for5 hours under a hydrogen atmosphere. The reaction solution was filtered,and the filtrate was concentrated under reduced pressure, and a mixedsolution of 2.27 g of the residue in 135 ml of DMF and 135 ml ofpyridine was stirred with 3.56 g (8.04 mmol) of BOP reagent at roomtemperature for 12 hours. The reaction solution was concentrated underreduced pressure, combined with water and extracted with ethyl acetate,and then the extract was successively washed with 10% aqueous citricacid solution and saturated aqueous sodium chloride solution. Theorganic layer was dried over magnesium sulfate, and then the solvent wasdistilled off under reduced pressure, and the residue was purified bysilica gel column chromatography (developing solvent; methylenechloride:methanol:aqueous ammonia=20:1:0.1) to give 1.14 g (60%) of(2S-(2S,13S))-2-tert-butoxycarbonylamino-N-(2-(3-tert-butyl-4-hydroxyphenylmethyl)-9-methyl-1,4,9-triaza-3,8,14-trioxocyclotetradec-13-yl)-3-(4-fluorophenyl)-N-methylpropionamide.

[0310] (6) Synthesis of(2S-(2S,13S))-2-amino-N-(2-(3-tert-butyl-4-hydroxyphenylmethyl)-9-methyl-1,4,9-triaza-3,8,14-trioxocyclotetradec-13-yl)-3-(4-fluorophenyl)-N-methylpropionamide

[0311] To a solution of 1.10 g (1.54 mmol) of(2S-(2S,13S))-2-tert-butoxycarbonylamino-N-(2-(3-tert-butyl-4-hydroxyphenylmethyl)-9-methyl-1,4,9-triaza-3,8,14-trioxocyclotetradec-13-yl)-3-(4-fluorophenyl)-N-methylpropionamidein 9 ml of methylene chloride was added 3 ml of TFA, and the mixture wasstirred at room temperature for 1 hour. The reaction solution wasdistilled off under reduced pressure, and the residue was neutralizedwith saturated aqueous NaHCO₃ solution, extracted with methylenechloride and the extract was washed with saturated aqueous sodiumchloride solution. The organic layer was dried over magnesium sulfate,and then the solvent was distilled off under reduced pressure, and theresidue was purified by silica gel column chromatography (developingsolvent; methylene chloride methanol:aqueous ammonia=13:1:0.1) to give609 mg (65%) of the title compound.

[0312] FAB-MS (M⁺+1): 612.

[0313]¹H-NMR (DMSO-d6-CDCl₃): δ 0.8-2.0 (6H, m), 1.29 (9H, s), 2.0-3.4(12H, m), 3.6-4.5 (3.5H, m), 4.9-5.1 (0.75H, m), 5.4-5.7 (0.75H, m),6.6-7.3 (10H, m), 7.6-7.8 (1H, m), 8.0-8.3 (1H, m), 8.9-9.2 (2H, m).

Example 10 Synthesis of(2S-(2S,13S))-2-amino-N-(2-(3-tert-butyl-4-hydroxyphenylmethyl)-1,4,8-triaza-3,7,14-trioxocyclotetradec-13-yl)-3-phenyl-N-methylpropionamide

[0314] (1) Synthesis of H-Tyr (3-tert-Bu)-β-Ala-O-tert-Bu

[0315] To a mixed solution of 3.5 g (13.9 mmol) of H-Tyr (3-tert-Bu)-OMeand 2.21 g (20.9 mmol) of sodium carbonate in 25 ml of 1,4-dioxane and25 ml of water was added 2.09 ml (14.6 mmol) of benzyloxycarbonylchloride with ice cooling, and the mixture was stirred at roomtemperature for 2 hours. The reaction solution was combined with water,extracted with ethyl ether, and then the extract was washed withsaturated aqueous sodium chloride solution. The organic layer was driedover anhydrous magnesium sulfate, and then the solvent was distilled offunder reduced pressure, and the resulting crude Z-Tyr (3-tert-Bu)-OMe in20 ml of 1,4-dioxane was stirred with a solution of 1.05 g (26.3 mmol)of sodium hydroxide in 10 ml of water at room temperature for 40minutes. The reaction solution was washed with ethyl ether, and then theaqueous layer was acidified with concentrated hydrochloric acid andextracted with methylene chloride. The extract was dried over anhydrousmagnesium sulfate, and then concentrated under reduced pressure, and1.88 g (13.9 mmol) of HOBt, 1.6 ml (13.9 mmol) of NMM and 2.93 g (15.3mmol) of WSCI were added to a solution of the resulting crude Z-Tyr(3-tert-Bu)-OH and 2.78 g (15.3 mmol) of H-β-Ala-O-tert-Bu hydrochloridein 40 ml of DMF with ice cooling, and the mixture was stirred overnightat room temperature. The reaction solution was combined with water, 10%aqueous citric acid solution and extracted with ethyl acetate, and thenthe extract was successively washed with saturated aqueous NaHCO₃solution and saturated aqueous sodium chloride solution. The extract wasdried over magnesium sulfate, and then the solvent was distilled offunder reduced pressure, and a solution of the resulting crude Z-Tyr(3-tert-Bu)-β-Ala-O-tert-Bu in 70 ml of methanol was stirred with 1.0 gof 20% palladium hydroxide on carbon for 1 hour under a hydrogenatmosphere. The reaction solution was filtered, and the filtrate wasconcentrated under reduced pressure to give 4.70 g (93%) of the titlecompound.

[0316]¹H-NMR (CDCl₃): δ 1.39 (9H, s), 1.45 (9H, s), 2.44 (2H, t, J=6.3Hz), 2.57 (1H, dd, J=9.4, 13.9 Hz), 3.17 (1H, dd, J=4.0, 13.9 Hz),3.47-3.56 (3H, m), 6.64 (1H, d, J=7.9 Hz), 6.90 (1H, dd, J=2.0, 7.9 Hz),7.08 (1H, d, J=2.0 Hz), 7.67 (1H, brt).

[0317] (2) Synthesis of Fmoc-N-Me-Lys (Boc)-Tyr(3-tert-Bu)-β-Ala-O-tert-Bu

[0318] To a solution of 3.97 g (10.9 mmol) of H-Tyr(3-tert-Bu)-β-Ala-O-tert-Bu and 5.26 g (10.9 mmol) of Fmoc-N-Me-Lys(Boc)-OH in 40 ml of DMF were added 1.47 g (10.9 mmol) of HOBt, 1.25 ml(10.9 mmol) of NMM and 2.09 g (10.9 mmol) of WSCI with ice cooling, andthe mixture was stirred overnight at room temperature. The reactionsolution was combined with water and 10% aqueous citric acid solutionand extracted with ethyl acetate, and then the extract was successivelywashed with saturated aqueous NaHCO₃ solution and saturated aqueoussodium chloride solution. The organic layer was dried over magnesiumsulfate, and then the solvent was distilled off under reduced pressure,and the residue was purified by silica gel column chromatography (usingethyl acetate:n-hexane=1:1 as eluent) to give 8.14 g (90%) of the titlecompound.

[0319]¹H-NMR (CDCl₃): δ 1.0-1.9 (6H, m), 1.34 (9H, s), 1.42 (9H, s),1.44 (9H, s), 2.3-2.4 (2H, m), 2.55 (3H, brs), 2.7-2.9 (1H, m), 3.0-3.1(3H, m), 3.4-3.5 (2H, m), 4.2-4.6 (6H, m), 6.45 (2H, brs), 6.75 (1H,brs), 7.00 (1H, s), 7.3-7.5 (5H, m), 7.58 (2H, d, J=6.3 Hz), 7.79 (2H,d, J=7.6 Hz).

[0320] (3) Synthesis of N-Me-Lys (Boc)-Tyr (3-tert-Bu)-β-Ala-O-tert-Bu

[0321] To a solution of 6.8 g (8.20 mmol) of Fmoc-N-Me-Lys (Boc)-Tyr(3-tert-Bu)-β-Ala-O-tert-Bu in 15 ml of methylene chloride was added 15ml of diethylamine, and the mixture was stirred at room temperature for95 minutes. The reaction solution was distilled off under reducedpressure, and the residue was purified by silica gel columnchromatography (using methylene chloride:methanol aqueousammonia=30:1:0.1 as eluent) to give 4.57 g (91%) of the title compound.

[0322]¹H-NMR (CDCl₃): δ 0.8-0.9 (2H, m), 1.0-1.7 (4H, m), 1.38 (9H, s),1.44 (9H, s), 1.48 (9H, s), 2.33 (3H, s), 2.43 (2H, t, J=5.9-6.3 Hz),2.7-3.1 (4H, m), 3.25 (1H, dd, J=4.6, 14.2 Hz), 3.49 (2H, dd, J=5.9-6.3,11.9-12.2 Hz), 4.5-4.7 (2H, m), 6.7-6.8 (2H, m), 6.86 (1H, d, J=6.6 Hz),7.03 (1H, s), 7.38 (1H, d, J=7.9 Hz).

[0323] (4) Synthesis of Z-Phe-N-Me-Lys (Boc)-Tyr(3-tert-Bu)-β-Ala-O-tert-Bu

[0324] To a solution of 4.57 g (7.53 mmol) of N-Me-Lys (Boc)-Tyr(3-tert-Bu)-β-Ala-O-tert-Bu, 2.93 g (9.79 mmol) of Z-Phe-OH and 2.69 g(10.5 mmol) of CMPI in 20 ml of THF was added 3.1 ml (22.6 mmol) of TEAwith ice cooling, and the mixture was stirred overnight at roomtemperature. The reaction solution was combined with water and extractedwith ethyl acetate, and then the extract was successively washed withsaturated aqueous NaHCO₃ solution and saturated aqueous sodium chloridesolution. The organic layer was dried over magnesium sulfate, and thenthe solvent was distilled off under reduced pressure, and the residuewas purified by silica gel column chromatography (developing solvent;ethyl acetate:hexane=3:1) to give 6.7 g (quant.) of the title compound.

[0325] (5) Synthesis of(2S-(2S,13S))-2-benzyloxycarbonylamino-N-(2-(3-tert-butyl-4-hydroxyphenylmethyl)-1,4,8-triaza-3,7,14-trioxocyclotetradec-13-yl)-3-phenyl-N-methylpropionamide

[0326] To a solution of 6.7 g (7.53 mmol) of Z-Phe-N-Me-Lys (Boc)-Tyr(3-tert-Bu)-β-Ala-O-tert-Bu in 40 ml of methylene chloride was added 40ml of TFA, and the mixture was stirred at room temperature for 70minutes. The reaction solution was concentrated under reduced pressure,and the residue was combined with toluene, and the solvent was distilledoff under reduced pressure. A mixed solution of the resulting crudeZ-Phe-N-Me-Lys-Tyr (3-tert-Bu)-β-Ala-OH in 380 ml of DMF and 380 ml ofpyridine was stirred with 9.99 g (22.6 mmol) of BOP reagent overnight atroom temperature. The reaction solution was concentrated under reducedpressure, and the residue was purified by silica gel columnchromatography (developing solvent; chloroform methanol:aqueousammonia=80-20:1:0.05). The residue was dissolved in ethyl acetate, andsuccessively washed with 10% aqueous citric acid solution, saturatedaqueous NaHCO₃ solution and saturated aqueous sodium chloride solution,and dried over anhydrous magnesium sulfate, and then the solvent wasdistilled off under reduced pressure to give 2.6 g (48%) of the titlecompound.

[0327] (6) Synthesis of(2S-(2S,13S))-2-amino-N-(2-(3-tert-butyl-4-hydroxyphenylmethyl)-1,4,8-triaza-3,7,14-trioxocyclotetradec-13-yl)-3-phenyl-N-methylpropionamide

[0328] To a solution of 1.25 g (1.79 mmol) of(2S-(2S,13S))-2-benzyloxycarbonylamino-N-(2-(3-tert-butyl-4-hydroxyphenylmethyl)-1,4,8-triaza-3,7,14-trioxocyclotetradec-13-yl)-3-phenyl-N-methylpropionamidein 35 ml of methanol was added 0.55 g of 10% palladium hydroxide oncarbon, and the mixture was stirred for 2.5 hours under a hydrogenatmosphere. The reaction solution was filtered, and the filtrate wasconcentrated under reduced pressure, and the residue was purified bysilica gel column chromatography (developing solvent;chloroform:methanol:aqueous ammonia=20:1:0.1) to give 1.48 g (73%) ofthe title compound.

[0329] FAB-MS (M⁺+1): 580

[0330]¹H-NMR (DMSO-d6-CDCl₃): δ 0.60-2.0 (6H, m), 1.29 (9H, s), 2.1-2.3(2H, m), 2.5-2.9 (4H, m), 2.66 and 2.60 (3H, s), 3.1-3.3 (2H, m),3.5-3.6 (2H, m), 3.71 and 3.92 (1H, t, J=6.3 Hz), 4.18 and 4.94 (1H, d,J=11.9 Hz), 4.4-4.6 (1H, m), 6.60-6.64 (1H, m), 6.72 and 6.75 (1H, dd,J=1.9, 8.0 Hz), 6.86 and 6.93 (1H, d, J=1.9 Hz), 7.05-7.29 (5H, m),7.6-7.8 (2H, m), 8.09 and 8.39 (1H, d, J=8.8-9.4), 8.9-9.1 (1H, s).

[0331] In the following test examples, representative compounds of thepresent invention were evaluated for motilin receptor antagonistactivity by pharmacological tests in order to demonstrate the utility ofcompounds of the present invention.

Test Example 1 Motilin Receptor Binding Test

[0332] Motilin receptor binding activities of compounds of the presentinvention were tested as follows [Vantrappen et al., Regul. Peptides,15, 143 (1986)].

[0333] A protein solution was prepared by homogenizing the duodenumisolated from a killed rabbit in 50 mM Tris after stripping the mucosa.The protein solution was incubated with 25 pM ¹²⁵I motilin for 2 hoursat 25° C., and then the radioactivity bound to the protein was measuredwith a γ-counter (COBRA II™, model 5005, Packard). The differencebetween the radioactivity of the protein solution incubated alone andthe radioactivity of the protein solution incubated with a large excessof motilin (10⁻⁷ M) was recorded as specific binding of motilin tomotilin receptors.

[0334] The concentration required to inhibit 50% of the specific bindingof motilin to motilin receptors (IC₅₀, nM) was determined for thecompounds synthesized in Examples 1-10 above. As a comparative example,the IC₅₀ of the compound of formula (5) below, i.e. a cyclic peptidederivative having motilin antagonist activity (the compound described inJP-A-7-138284) was also determined. The results are shown in Table B-1.

Test Example 2 Effect on the Contraction of a Sample of the LongitudinalDuodenal Muscle Extracted from Rabbit

[0335] The effect of compounds of the present invention onmotilin-induced contraction of a sample of the longitudinal muscle ofthe duodenum extracted from a rabbit was tested as follows.

[0336] A duodenal specimen (5×15 mm) extracted from a killed rabbit wassuspended in the direction of the longitudinal muscle in an incubator at28° C. filled with Krebs solution (10 ml organ bath). A mixed gas (95%O₂, 5% CO₂) was continuously passed through the Krebs solution and thecontraction of the duodenal specimen was isotonically recorded (under aload of 1 g) via an isotonic transducer (ME-3407, ME Commercial, Tokyo,Japan). The extent of contraction of the duodenal specimen was expressedas a percentage to the contraction induced by acetylcholine at aconcentration of 10⁻⁴ M.

[0337] The effect of the compounds synthesized in Examples 1-8 and 10 onthe concentration-dependent contraction induced by motilin added intothe incubator was determined to calculate pA₂ (see “Drug receptors”edited by Takayanagi, published by Nanzando). The results are shown inTable B-1. As a comparative example, the results of the compound offormula (5) are also shown. TABLE B-1 Motilin receptor Contractionbinding test inhibition test Example No. IC₅₀ (nM) pA₂ 1 1.2 9.0 2 1.48.9 3 0.52 8.2 4 0.78 9.0 5 1.4 9.9 6 0.87 8.5 7 4.5 8.0 8 0.84 8.2 95.3 10  2.2 7.9 Comparative 10 7.2 example

[0338] Table B-1 shows that the compounds of the present invention haveexcellent motilin receptor antagonist activity as compared with thecompound of the comparative example.

INDUSTRIAL APPLICABILITY

[0339] Compounds of the present invention have motilin receptorantagonist activity or the like and are useful as pharmaceuticals suchas pharmaceuticals for treating irritable bowel syndrome.

1. A compound of general formula (1):

wherein R₁ represents an optionally substituted phenyl group or anoptionally substituted heterocycle; R₂ represents a hydrogen atom or anoptionally substituted amino group; R₃ represents a hydrogen atom, amethyl group or an ethyl group; R₄ represents a hydrogen atom, a methylgroup or an ethyl group; R₅ represents a hydrogen atom, a methyl groupor an ethyl group; R₆ represents a hydrogen atom, a methyl group, anethyl group or —CO—R₇; R₇ represents a hydrogen atom or a straight orbranched alkyl group having 1-3 carbon atoms; V represents a carbonylgroup or a methylene group; W represents a carbonyl group or a methylenegroup: X represents a carbonyl group or a methylene group; Y representsa carbonyl group or a methylene group; Z represents a carbonyl group ora methylene group; m represents a number of 0-2; and n represents anumber of 0-3; except for the case where R₁ represents a phenyl group,R₂ represents an amino group, all of R₃, R₄, R₅ and R₆ represent ahydrogen atom, V represents a methylene group, all of W, X, Y and Zrepresent a carbonyl group, and both m and n represent 1; or a hydrateor a pharmaceutically acceptable salt thereof.
 2. The compound ofgeneral formula (1) of claim 1 wherein X is a methylene group when R₁ isa phenyl group and R₃ is a hydrogen atom, or a hydrate or apharmaceutically acceptable salt thereof.
 3. The compound of generalformula (1) of claim 1 or 2 wherein R₁ is a phenyl group or ahalogen-substituted phenyl group, or a hydrate or a pharmaceuticallyacceptable salt thereof.
 4. The compound of general formula (1) of anyone of claims 1 to 3 wherein R₂ is an amino group, or a hydrate or apharmaceutically acceptable salt thereof.
 5. The compound of generalformula (1) of any one of claims 1 to 4 wherein R₃ is a hydrogen atom ora methyl group, or a hydrate or a pharmaceutically acceptable saltthereof.
 6. The compound of general formula (1) of any one of claims 1to 5 wherein R₄ is a hydrogen atom or a methyl group, or a hydrate or apharmaceutically acceptable salt thereof.
 7. The compound of generalformula (1) of any one of claims 1 to 6 wherein R₅ is a hydrogen atom ora methyl group, or a hydrate or a pharmaceutically acceptable saltthereof.
 8. The compound of general formula (1) of any one of claims 1to 7 wherein R₆ is a hydrogen atom, a methyl group or an acetyl group,or a hydrate or a pharmaceutically acceptable salt thereof.
 9. Thecompound of general formula (1) of any one of claims 1 to 8 wherein V isa methylene group, or a hydrate or a pharmaceutically acceptable saltthereof.
 10. The compound of general formula (1) of any one of claims 1to 9 wherein Y is a carbonyl group, or a hydrate or a pharmaceuticallyacceptable salt thereof.
 11. The compound of general formula (1) of anyone of claims 1 to 10 wherein Z is a carbonyl group, or a hydrate or apharmaceutically acceptable salt thereof.
 12. The compound of generalformula (1) of claim 1 wherein R₁ is a phenyl group or ahalogen-substituted phenyl group, R₂ is an amino group, R₃ is a hydrogenatom or a methyl group, R₄ is a hydrogen atom or a methyl group, R₅ is ahydrogen atom or a methyl group, R₆ is a hydrogen atom, a methyl groupor an acetyl group, V is a methylene group, W is a carbonyl group or amethylene group, X is a carbonyl group or a methylene group, Y is acarbonyl group and Z is a carbonyl group, or a hydrate or apharmaceutically acceptable salt thereof.
 13. The compound of claim 1selected from the group consisting of(2S-(2S,13S))-2-amino-N-(2-(3-tert-butyl-4-hydroxyphenylmethyl)-1,4,8-triaza-3,7,14-trioxocyclotetradec-13-yl)-3-(4-fluorophenyl)-N-methylpropionamide,(2S-(2S,13S))-2-amino-N-(2-(3-tert-butyl-4-hydroxyphenylmethyl)-1-methyl-1,4,8-triaza-3,7,14-trioxocyclotetradec-13-yl)-3-(4-fluorophenyl)-N-methylpropionamide,(2S-(2S,12S))-2-amino-N-(2-(3-tert-butyl-4-hydroxyphenylmethyl)-1,4,8-triaza-3,7,13-trioxocyclotridec-12-yl)-3-(4-fluorophenyl)-N-methylpropionamide,(2S-(2S,13S))-2-amino-N-(2-(3-tert-butyl-4-hydroxyphenylmethyl)-4-methyl-1,4,8-triaza-3,7,14-trioxocyclotetradec-13-yl)-3-(4-fluorophenyl)-N-methylpropionamide,(2S-(2S,13S))-2-amino-N-(2-(3-tert-butyl-4-hydroxyphenylmethyl)-8-methyl-1,4,8-triaza-3,7,14-trioxocyclotetradec-13-yl)-3-(4-fluorophenyl)-N-methylpropionamide,(2S-(2S,14S))-2-amino-N-(2-(3-tert-butyl-4-hydroxyphenylmethyl)-1,4,9-triaza-3,8,15-trioxocyclopentadec-14-yl)-3-(4-fluorophenyl)-N-methylpropionamide,(2S,14S)-13-(2S-2-amino-3-(4-fluorophenyl)propylamino)-2-(3-tert-butyl-4-hydroxyphenylmethyl)-1,4,8-triaza-3,7,14-trioxocyclotetradecane,(2S-(2S,13S))—N-(8-acetyl-2-(3-tert-butyl-4-hydroxyphenylmethyl)-N-methyl-1,4,8-triaza-3,14-dioxocyclotetradec-13-yl)-2-amino-3-(4-fluorophenyl)propionamide,(2S-(2S,13S))-2-amino-N-(2-(3-tert-butyl-4-hydroxyphenylmethyl)-9,N-dimethyl-1,4,9-triaza-3,8,14-trioxocyclotetradec-13-yl)-3-(4-fluorophenyl)propionamide,(2S-(2S,13S))-2-amino-N-(2-(3-tert-butyl-4-hydroxyphenylmethyl)-1,4,8-triaza-3,7,14-trioxocyclotetradec-13-yl)-3-phenyl-N-methylpropionamide,or a hydrate or a pharmaceutically acceptable salt thereof.
 14. Apharmaceutical comprising the compound of any one of claims 1 to 13 or ahydrate or a pharmaceutically acceptable salt thereof as an activeingredient.
 15. A motilin receptor antagonist comprising the compound ofany one of claims 1 to 13 or a hydrate or a pharmaceutically acceptablesalt thereof.
 16. A gastrointestinal motility inhibitor comprising thecompound of any one of claims 1 to 13 or a hydrate or a pharmaceuticallyacceptable salt thereof as an active ingredient.
 17. A pharmaceuticalfor treating hypermotilinemia comprising the compound of any one ofclaims 1 to 13 or a hydrate or a pharmaceutically acceptable saltthereof as an active ingredient.
 18. A compound of general formula (2):

wherein R₃ represents a hydrogen atom, a methyl group or an ethyl group;R₄ represents a hydrogen atom, a methyl group or an ethyl group; R₅represents a hydrogen atom, a methyl group or an ethyl group; R₆represents a hydrogen atom, a methyl group, an ethyl group or —CO—R₇; mrepresents a number of 0-2; n represents a number of 0-3; P₂ representsa hydrogen atom or a protective group for a phenolic hydroxyl group; P₃represents a hydrogen atom or a protective group for a carboxyl group;P₄ represents a hydrogen atom or a protective group for an amino group;and P₅ represents a hydrogen atom or a protective group for an aminogroup; or a hydrate or a pharmaceutically acceptable salt thereof.
 19. Acompound of general formula (3):

wherein R₁ represents an optionally substituted phenyl group or anoptionally substituted heterocycle; R₃ represents a hydrogen atom, amethyl group or an ethyl group; R₄ represents a hydrogen atom, a methylgroup or an ethyl group; R₅ represents a hydrogen atom, a methyl groupor an ethyl group; m represents a number of 0-2; n represents a numberof 0-3; P₂ represents a hydrogen atom or a protective group for aphenolic hydroxyl group; P₃ represents a hydrogen atom or a protectivegroup for a carboxyl group; P₅ represents a hydrogen atom or aprotective group for an amino group; and R₂′ represents a hydrogen atomor an optionally substituted protected amino group; or a hydrate or apharmaceutically acceptable salt thereof.
 20. A compound of generalformula (4):

wherein R₁ represents an optionally substituted phenyl group or anoptionally substituted heterocycle; R₃ represents a hydrogen atom, amethyl group or an ethyl group; R₄ represents a hydrogen atom, a methylgroup or an ethyl group; R₅ represents a hydrogen atom, a methyl groupor an ethyl group; R₂′ represents a hydrogen atom or an optionallysubstituted protected amino group; m represents a number of 0-2; nrepresents a number of 0-3; P₆ represents a hydrogen atom or aprotective group for a carboxyl group; and P₇ represents a hydrogen atomor a protective group for an amino group; or a hydrate or apharmaceutically acceptable salt thereof.
 21. A method for treatinghypermotilinemia comprising the step of administering a therapeuticallyeffective amount of the compound of any one of claims 1 to 13 or ahydrate or a pharmaceutically acceptable salt thereof to a patient inneed of such treatment.